Bicyclic Compounds and their Uses

ABSTRACT

The present invention provides a compound, or a pharmaceutically acceptable salt thereof, of formula (I):wherein R1, R2, R3, R4, R5, R26, R27, y, R, M, W, L, V, T, Y, J, K and A are as described herein, therapeutic uses of said compounds, uses of said compounds as research chemicals, a pharmaceutical composition and combinations comprising said compounds, and methods for manufacturing the compounds of the invention.

FIELD OF INVENTION

The invention provides bicyclic compounds, such as7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl) compounds, and analoguesand derivatives thereof, the use thereof for inhibiting Werner SyndromeRecQ DNA helicase (WRN) and methods of treating disease using saidcompounds, in particular the use in treating cancer, and in particularthe treatment of cancer characterized as microsatellite instability-high(MSI-H) or mismatch repair deficient (dMMR), including colorectal,gastric and endometrial cancer. The invention also provides the use ofsaid compounds as research chemicals, intermediate compounds,combinations, processes and formulations.

BACKGROUND OF THE INVENTION

Loss of DNA mismatch repair is a common initiating event in cancerdevelopment occurring in 10-30% of colorectal, endometrial, ovarian andgastric cancers (Aaltonen, L. A. et al. Clues to the pathogenesis offamilial colorectal cancer, Science 260, 812-816 (1993), Bonneville R etal., Landscape of Microsatellite Instability Across 39 Cancer Types. JCOPrecis Oncol. 1: PO.17.00073 (2017)). Cancers that have lost competencein mismatch repair (MMR) have a high mutational burden, and frequentdeletion and insertion events in repetitive DNA tracts, a phenotypeknown as microsatellite instability (MSI). While progress has been madein the treatment of microsatellite instability high (MSI-H) cancers, andthe demonstration that pembrolizumab (anti-PD1) treatment led tosignificantly longer progression-free survival than chemotherapy whenreceived as first-line therapy for MSI-H-dMMR metastatic colorectalcancer resulted in the recent approval of pembrolizumab as first-linetreatment of these cancers, there is still a significant unmet medicalneed in CRC and other MSI-H indications (André T., et al. Pembrolizumabin Microsatellite-Instability-High Advanced Colorectal Cancer. N Engl JMed 383(23):2207-2218 (2020)). Several large-scale functional genomicsscreens across large panels of cell lines, including Novartis with 398cell lines from the Cancer Cell Line Encyclopedia (CCLE) (McDonald E. R.et al., Project DRIVE: A Compendium of Cancer Dependencies and SyntheticLethal Relationships Uncovered by Large-Scale, Deep RNAi Screening. Cell170(3):577-592 (2017)), have identified the Werner Syndrome RecQhelicase (WRN) as being selectively required for the survival of celllines with defective mismatch repair that have become MSI-H (Behan, F.M. et al. Prioritization of cancer therapeutic targets using CRISPR-Cas9screens. Nature 568, 511-516 (2019), Chan, E. M. et al. WRN helicase isa synthetic lethal target in microsatellite unstable cancers. Nature568, 551-556 (2019). Kategaya, L., Perumal, S. K., Hager, J. H. &Belmont, L. D. Werner syndrome helicase is required for the survival ofcancer cells with microsatellite instability. iScience 13, 488-497(2019), Lieb, S. et al. Werner syndrome helicase is a selectivevulnerability of microsatellite instability-high tumor cells. eLife 8,e43333 (2019)). WRN is synthetic lethal with MSI cancers. Depletion ofWRN leads to anti-proliferative effects and results in activation ofmultiple DNA damage signaling markers, induction of cell cycle arrestand apoptosis in MMR cancer models but not cancer cells with an intactMMR pathway. These findings indicate that WRN provides a DNA repair andmaintenance function that is essential for cell survival in MSI cancers.Recently, the mechanism of WRN dependence has been elucidated. It hasbeen shown that dinucleotide TA repeats are selectively unstable in MSIcells and undergo large scale expansions. These expanded TA repeats formsecondary DNA structures that require the WRN helicase for unwinding(van Wietmarschen, N. et al. Repeat expansions confer WRN dependence inmicrosatellite-unstable cancers. Nature 586, 292-298, 2020). In theabsence of WRN (or upon WRN helicase inhibition), expanded TA repeats inMSI cells are subject to nuclease cleavage and chromosome breakage.Thus, inhibiting the WRN helicase is an attractive strategy for thetreatment of mismatch repair defective cancers.

SUMMARY OF THE INVENTION

There remains a need for new treatments and therapies for the treatmentof cancer, and in particular cancers characterized as microsatelliteinstability-high (MSI-H) or mismatch repair deficient (dMMR), includingcolorectal, gastric or endometrial cancer. The invention providescompounds, pharmaceutically acceptable salts thereof, pharmaceuticalcompositions thereof and combinations thereof, said compounds beinginhibitors of Werner Syndrome RecQ DNA Helicase (WRN). The inventionfurther provides methods of treating, preventing, or ameliorating adisease or condition, comprising administering to a subject in needthereof an effective amount of a WRN inhibitor. The invention alsoprovides compounds, pharmaceutically acceptable salts thereof,pharmaceutical compositions thereof and combinations thereof, saidcompounds being useful for the treatment of cancer, in particularcancers characterized as microsatellite instability-high (MSI-H) ormismatch repair deficient (dMMR). Also provided are compounds that bindto, and/or inhibit WRN, and are therefore useful as research chemicals,e.g. as a chemical probe, and as tool compounds. Various embodiments ofthe invention are described herein.

Within certain aspects, provided herein is a compound of formula (I) ora pharmaceutically acceptable salt thereof:

wherein

R, M, W, L, V and T are independently selected from C, CH and N,

to form subformulae 1a, 1b, 1c, 1d, 1e and 1f:

A is a linker selected from —C(O)—, —S(O)—, —S(O)₂—, and

Y is N, C or CH;

y is 0, 1, 2, 3 or 4;

Y

means Y is linked via a single bond to the adjacent carbon atom when Yis CH, or Y is linked via a double bond to the adjacent atom when Y isC, and when Y

is a single bond, Y is carbon unsubstituted or substituted by OH or F;

when Y is N, Y

is a single bond;

K

means K is linked via a single or double bond to the adjacent atom;

wherein:

-   -   when K        is a double bond, Y        is a single bond, K is CH, J is C, and A is a linker selected        from —C(O)—, —S(O)—, —S(O)₂—, and

or

-   -   when K        is a single bond, K is selected from —CH₂—, —CH₂CH₂—, —NH— and a        bond (to form a 5-membered ring

J is N, and A is a linker selected from —C(O)—, —S(O)—, —S(O)₂—, and

or

-   -   when K        is a single bond, K is —CH₂—, J is CH, and A is a linker        selected from —S(O)—, —S(O)₂—, and

R₅ is independently selected from:

-   -   —(C₁-C₄)alkyl,    -   —(C₃-C₅)cycloalkyl,    -   and wherein two R₅ substituents on the same ring carbon atom may        join, together with the carbon atom to which they are attached,        to form a (C₃-C₄)cycloalkyl spiro ring or a 3 or 4-membered        heterocyclyl spiro ring, wherein said heterocyclyl spiro ring        contains ring carbon ring atoms and one ring heteroatom selected        from O, N and S,    -   when K        is a carbon-nitrogen single bond, a R₅ substituent on K and on        the adjacent carbon atom may join to form ring C:

-   -   wherein ring C is a fused (C₃-C₆)cycloalkyl ring, a fused        (C₃-C₆)heterocyclyl ring or a fused phenyl ring, wherein said        fused (C₃-C₆)heterocyclyl ring contains ring carbon atoms and        one ring heteroatom selected from O, N and S,    -   when K        J is a carbon-carbon single bond, Y is N and Y        is a single bond, and    -   A is a linker selected from —S(O)—, —S(O)₂—, and

a R₅ substituent on K and on the adjacent carbon atom may join to formring C:

-   -   and wherein when K is —CH₂— and J is N, two R₅ substituents may        join to form a (C₁-C₃)alkylene bridge or a heteroalkylene        bridge, wherein said heteroalkylene bridge is one heteroatom        selected from N and O, or is —CH₂—O—CH₂—;

and wherein one or more H atoms on the ring:

may be replaced by deuterium;

R₁ is:

cycloalkenyl, wherein said cycloalkenyl is a partially unsaturatedmonocyclic ring containing 5 or 6 ring carbon atoms, and saidcycloalkenyl is unsubstituted or substituted by 1, 2, 3 or 4, preferably1 or 2, R₃₃, wherein R₃₃ is halo, and wherein said cycloalkenyl orhalo-substituted cycloalkenyl is substituted by 0, 1 or 2 R₁₅substituents,

or R₁ is heterocyclyl, wherein said heterocyclyl is a 5 or 6 memberedfully saturated or partially unsaturated group comprising ring carbonatoms and 1 or 2 ring heteroatoms independently selected from N, O andS, and wherein said heterocyclyl is unbridged or bridged, and saidbridge is 1 or 2 carbon atoms, wherein said heterocyclyl isunsubstituted or substituted by 1, 2, 3 or 4, preferably 1 or 2, R₃₃,wherein R₃₃ is halo, and wherein said heterocyclyl or halo-substitutedheterocyclyl is substituted by 0, 1 or 2 substituents independentlyselected from R₁₅, R₁₆, R₁₇, R₁₈, R₁₉, R₂₀, R₂₂ and R₂₃,

or said heterocyclyl or halo-substituted heterocyclyl is fused to acyclopropyl ring, wherein said cyclopropyl ring is unsubstituted orsubstituted by 1, 2 or 3 F,

or said heterocyclyl or halo-substituted heterocyclyl has 2substitutents at the same ring carbon atom which join to form acyclopropyl spiro ring,

or said heterocyclyl or halo-substituted heterocyclyl is fused with a(C₃-C₅)heterocycloalkyl ring, wherein said (C₃-C₅)heterocycloalkyl ringcontains ring carbon atoms and 1 ring 0 atom;

or R₁ is heteroaryl, wherein said heteroaryl is a 5 or 6 membered fullyunsaturated monocyclic group comprising ring carbon atoms and 1, 2, 3 or4 ring heteroatoms independently selected from N, O and S, preferably 1or 2 ring heteroatoms, wherein the total number of ring S atoms does notexceed 1, and the total number of ring O atoms does not exceed 1,wherein said heteroaryl is unsubstituted or substituted by 1, 2 or 3substituents independently selected from R₂₁ and R₃₀, wherein R₂₁ andR₃₀ are independently selected from halo and (C₁-C₄)alkyl, wherein said(C₁-C₄)alkyl is unsubstituted or substituted by 1, 2 or 3 halo,

or R₁ is phenyl, wherein said phenyl is unsubstituted or substituted by1, 2, 3 or 4, preferably 1 or 2, R₃₃, wherein R₃₃ is halo, and whereinsaid phenyl or halo-substituted phenyl is substituted by 0, 1 or 2 R₁₅substituents,

or R₁ is (C₂-C₄)alkynyl or (C₂-C₄)alkenyl, wherein said (C₂-C₄)alkynyland (C₂-C₄)alkenyl are unsubstituted or substituted by(C₁-C₄)alkyl-O—C(O)—, or morpholinyl;

each R₁₅, R₁₆, R₁₇, R₁₈, R₁₉, R₂₀, R₂₂ and R₂₃ is independently selectedfrom:

-   -   halo    -   (C₁-C₄)alkyl-O— unsubstituted or substituted by 1, 2 or 3 halo;    -   (C₁-C₄)alkyl unsubstituted or substituted by OH, —O—(C₁-C₂)alkyl        or 1, 2 or 3 halo,    -   HOC(O)—(CH₂)_(n)—,    -   H₃C—C(O)(CH₂)_(n)—,    -   (C₁-C₄)alkyl-O—C(O)(CH₂)_(n),    -   ═O    -   azetidinyl or pyrrolidinyl, wherein said azetidinyl and        pyrrolidinyl are linked to the rest of the molecule via the N        atom, and are each unsubstituted or substituted by 1 or 2 F,    -   R₂₅(R₂₄)N—, wherein R₂₄ is H or (C₁-C₄)alkyl unsubstituted or        substituted by 1, 2 or 3 halo, R₂₅ is H or (C₁-C₄)alkyl        unsubstituted or substituted by 1, 2 or 3 halo,    -   OH

wherein n is 0, 1 or 2,

R₂₆ is CH₃, H or deuterium;

R₂₇ is CH₃, H or deuterium;

or R₂₆ and R₂₇ join, together with the carbon atom to which they areattached, to form a cyclopropyl ring;

R₂ is the moiety:

R₆ is selected from:

-   -   H,    -   halo,    -   (C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo,    -   (C₃-C₅)cycloalkyl unsubstituted or substituted by 1, 2 or 3        halo,    -   —O—(C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo,    -   OH, and    -   CN;

R₈ is selected from H, halo, and (C₁-C₄)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo,

R₉ is selected from H, O—CH₃, OH, CN, CH₃ and halo;

R₂₃ is selected from:

-   -   SF₅,    -   H,    -   —C(O)H,    -   halo,    -   (C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo,    -   (C₁-C₄)alkynyl,    -   (C₁-C₄)alkenyl,    -   (C₃-C₅)cycloalkyl unsubstituted or substituted by 1, 2 or 3        halo, and    -   OCF₃;

X is selected from C—R₇ and N, wherein R₇ is H or halo, or R₇ can join,together with R₂₈ or R₆, and the atoms to which they are attached, toform a fused (C₄-C₆)cycloalkyl ring, wherein said fused(C₄-C₆)cycloalkyl ring is unsubstituted or substituted by 1, 2 or 3halo,

or

R₂ is selected from:

wherein

R₃₁ is selected from H, halo and CH₃,

R₃₂ is selected from H, halo and CH₃,

R₃ is:

-   -   cyclopropyl,    -   O—CH₃,    -   N(CH₃)₂,    -   S—CH₃,    -   (C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3        substituents independently selected from halo and OH;

R₄ is selected from:

wherein

R₁₀, R₁₁, R₁₂, R₁₃ and R₁₄ are independently selected from:

-   -   H,    -   halo,    -   (C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo        substituents,    -   (C₁-C₂)alkyl substituted by —O—(C₁-C₂)alkyl or OH,    -   —S—(C₁-C₃)alkyl,    -   O—(C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo        substituents,    -   OH,    -   (C₃-C₅)cycloalkyl, wherein said (C₃-C₅)cycloalkyl is        unsubstituted or substituted by 1 or 2 halo,    -   —O—(C₃-C₅)cycloalkyl,    -   —NR₃₄R₃₅ wherein R₃₄ and R₃₅ are independently selected from:        -   H,        -   (C₁-C₄)alkyl, wherein said (C₁-C₄)alkyl is unsubstituted or            substituted by OH or —O(C₁-C₂)alkyl,        -   and wherein R₃₄ and R₃₅ can join, together with the atom to            which they are attached, to form an azetidine, pyrrolidinyl            or piperidine ring, wherein said azetidine, pyrrolidinyl and            piperidine are unsubstituted or substituted with CH₃;    -   CN,    -   —(C₂-C₄)alkenyl,    -   —(C₂-C₄)alkynyl,    -   —C(O)H, and    -   —C(O)(C₁-C₄)alkyl;

and

* indicates a point of attachment.

In another aspect, the invention provides a pharmaceutical compositioncomprising a compound of formula (I) of the present invention and one ormore pharmaceutically acceptable carriers.

In another aspect, the invention provides a combination, in particular apharmaceutical combination, comprising a compound of formula (I) of thepresent invention and one or more therapeutically active agents.

In another aspect, the invention provides a compound of formula (I) ofthe present invention for use as a medicament, in particular for thetreatment of a disorder or disease which can be treated by WRNinhibition.

In another aspect, the invention provides a compound of formula (I) ofthe present invention for use in the treatment of cancer, particularlywherein the cancer is characterized as microsatellite instability-high(MSI-H) or mismatch repair deficient (dMMR).

In another aspect, the invention provides a method of treating adisorder or disease which can be treated by WRN inhibition in a subject,comprising administering to the subject a therapeutically effectiveamount of a compound of formula (I) of the present invention.

In another aspect, the invention provides a method of treating cancer ina subject, more particularly wherein the cancer is characterized asmicrosatellite instability-high (MSI-H) or mismatch repair deficient(dMMR), comprising administering to the subject a therapeuticallyeffective amount of a compound of formula (I) of the present invention.

In another aspect, the invention provides the use of a compound offormula (I) of the present invention in the manufacture of a medicamentfor the treatment of a disorder or disease which can be treated by WRNinhibition.

In another aspect, the invention provides a compound of formula (I) ofthe present invention for use as a research chemical, for example as achemical probe or as a tool compound.

In another aspect, the invention provides a solid form, process orintermediate as described herein.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a X-ray powder diffractogram for example 42.

FIG. 2 shows a X-ray powder diffractogram for example 86.

FIG. 3 shows a X-ray powder diffractogram for example 47.

FIG. 4 shows a X-ray powder diffractogram for example 57.

FIG. 5 shows a X-ray powder diffractogram for example 96.

FIG. 6 shows efficacy and tolerability of Example 58 after once daily(qd) administration in female nude mice bearing SW48 xenografts

FIG. 7 shows a X-ray powder diffractogram for example 42.

FIG. 8 shows the colony formation assay data for the compound of example42, using MSI and MSS cells.

FIG. 9 shows efficacy of Example 42 after administration in female nudemice bearing SW48 xenografts

FIG. 10 shows efficacy of Example 96 after administration in female nudemice bearing SW48 xenografts

FIG. 11 shows efficacy of Example 57 after administration in female nudemice bearing SW48 xenografts.

DETAILED DESCRIPTION

The invention therefore provides a compound of formula (I):

wherein R₁, R₂, R₃, R₄, R₅, R₂₆, R₂₇, y, R, M, W, L, V, T, Y, J, K and Aare as described in the Summary of the Invention, supra.

Unless specified otherwise, the term “compounds of the presentinvention” or “compound of the present invention” or “a compound offormula (I)”, refers to a compound or compounds of formula (I),subformulae thereof, exemplified compounds, and salts thereof, as wellas all zwitterions, stereoisomers (including diastereoisomers andenantiomers), rotamers, tautomers and isotopically labeled compounds(including deuterium substitutions), as well as inherently formedmoieties, and combinations or mixtures of the above-mentioned aspectsthereof.

Various (enumerated) embodiments of the invention are described herein.It will be recognized that features specified in each embodiment may becombined with other specified features to provide further embodiments ofthe present invention.

Embodiment 1. A compound of formula (I) or a pharmaceutically acceptablesalt thereof, as described above.

Embodiment 2: A compound of formula (I) or a pharmaceutically acceptablesalt thereof, according to Embodiment 1, wherein when R₁ is a ring,then:

-   -   each R₁ ring atom adjacent to the R₁ ring atom to which said R₁        ring is joined to the remainder of the molecule, is        independently unsubstituted or substituted by halo only, in        particular, independently unsubstituted or substituted with one        F substituent, and preferably,    -   said R₁ ring is linked to the remainder of the molecule via a R₁        ring nitrogen atom, or a R₁ ring carbon atom which is        double-bonded to an adjacent ring atom.

Embodiment 3. A compound of formula (I) or a pharmaceutically acceptablesalt thereof, according to Embodiments 1 or 2, wherein R₁ is:

cycloalkenyl, wherein said cycloalkenyl is a partially unsaturatedmonocyclic ring containing 5 or 6 ring carbon atoms, and saidcycloalkenyl is unsubstituted or substituted by 1, 2, 3 or 4, preferably1 or 2, R₃₃, wherein R₃₃ is halo, and wherein said cycloalkenyl orhalo-substituted cycloalkenyl is substituted by 0, 1 or 2 R₁₅substituents,

or R₁ is heterocyclyl, wherein said heterocyclyl is a 5 or 6 memberedfully saturated or partially unsaturated group comprising ring carbonatoms and 1 or 2 ring heteroatoms independently selected from N, O andS, and wherein said heterocyclyl is unbridged or bridged, and saidbridge is 1 or 2 carbon atoms, wherein said heterocyclyl isunsubstituted or substituted by 1, 2, 3 or 4, preferably 1 or 2, R₃₃,wherein R₃₃ is halo, and wherein said heterocyclyl or halo-substitutedheterocyclyl is substituted by 0, 1 or 2 substituents independentlyselected from R₁₅, R₁₆, R₁₇, R₁₈, R₁₉, R₂₀, R₂₂ and R₂₃,

or R₁ is heteroaryl, wherein said heteroaryl is a 5 or 6 membered fullyunsaturated monocyclic group comprising ring carbon atoms and 1, 2, 3 or4 ring heteroatoms independently selected from N, O and S, preferably 1or 2 ring heteroatoms, wherein the total number of ring S atoms does notexceed 1, and the total number of ring O atoms does not exceed 1,wherein said heteroaryl is unsubstituted or substituted by 1, 2 or 3substituents independently selected from R₂₁ and R₃₀, wherein R₂₁ andR₃₀ are independently selected from halo and (C₁-C₄)alkyl, wherein said(C₁-C₄)alkyl is unsubstituted or substituted by 1, 2 or 3 halo,

and each R₁₅, R₁₆, R₁₇, R₁₈, R₁₉, R₂₀, R₂₂ and R₂₃ is independentlyselected from:

-   -   halo    -   (C₁-C₄)alkyl-O— unsubstituted or substituted by 1, 2 or 3 halo;    -   (C₁-C₄)alkyl unsubstituted or substituted by OH, —O—(C₁-C₂)alkyl        or 1, 2 or 3 halo,    -   HOC(O)—(CH₂)_(n)—,    -   H₃C—C(O)(CH₂)_(n)—,    -   (C₁-C₄)alkyl-O—C(O)(CH₂)_(n),    -   ═O    -   azetidinyl or pyrrolidinyl, wherein said azetidinyl and        pyrrolidinyl are linked to the rest of the molecule via the N        atom, and are each unsubstituted or substituted by 1 or 2 F,    -   R₂₅(R₂₄)N—, wherein R₂₄ is H or (C₁-C₄)alkyl unsubstituted or        substituted by 1, 2 or 3 halo, R₂₅ is H or (C₁-C₄)alkyl        unsubstituted or substituted by 1, 2 or 3 halo,    -   OH

wherein n is 0, 1 or 2,

Embodiment 4. A compound of formula (I) or a pharmaceutically acceptablesalt thereof, according to any of Embodiments 1, 2 or 3, wherein R₁ is:

cycloalkenyl, wherein said cycloalkenyl is a partially unsaturatedmonocyclic ring containing 5 or 6 ring carbon atoms, and saidcycloalkenyl is unsubstituted or substituted by 1 or 2 R₃₃, wherein R₃₃is halo, preferably F, and wherein said cycloalkenyl or halo-substitutedcycloalkenyl is substituted by 0 or 1 R₁₅ substituents, preferably 1substituent, wherein R₁₅ is selected from:

-   -   a) (C₁-C₂)alkyl-O— unsubstituted or substituted by 1, 2 or 3        halo;    -   b) (C₁-C₂)alkyl unsubstituted or substituted by 1, 2 or 3 halo,    -   c) HOC(O)—(CH₂)_(n)—,    -   d) H₃C—C(O)(CH₂)_(n)—,    -   e) H₃C—O—C(O)(CH₂)_(n),    -   f) ═O, and    -   g) R₂₅(R₂₄)N—, H, wherein R₂₄ is H or (C₁-C₂)alkyl unsubstituted        or substituted by 1, 2 or 3 halo, R₂₅ is H or (C₁-C₂)alkyl        unsubstituted or substituted by 1, 2 or 3 halo,

n is 0 or 1,

wherein

-   -   the R₁₅ substituent a) to g) of said cycloalkenyl or        halo-substituted cycloalkenyl is not present on the ring atoms        adjacent to the ring atom to which the cycloalkenyl or        halo-substituted cycloalkenyl is joined to the remainder of the        molecule, and preferably, said cycloalkenyl or halo-substituted        cycloalkenyl is a 6 membered ring, with 1 R₁₅ substituent in the        ring para position relative to the remainder of the molecule;        and    -   said cycloalkenyl or halo-substituted cycloalkenyl is linked to        the remainder of the compound via a R₁ ring carbon atom which is        double bonded to an adjacent R₁ ring carbon atom;

or R₁ is heterocyclyl, wherein said heterocyclyl is a 5 or 6 memberedfully saturated or partially unsaturated group comprising ring carbonatoms and 1 or 2 ring heteroatoms independently selected from N, NH, Oand S, and wherein said heterocyclyl is unbridged or bridged, and saidbridge is 1 or 2 carbon atoms, wherein said heterocyclyl isunsubstituted or substituted by 1 or 2 R₃₃, wherein R₃₃ halo, ispreferably F, and wherein said heterocyclyl or halo-substitutedheterocyclyl is substituted by 0 or 1 substituents independentlyselected from R₁₅, R₁₆, R₁₇, R₁₈, R₁₉, R₂₀, R₂₂ and R₂₃, wherein saidR₁₅, R₁₆, R₁₇, R₁₈, R₁₉, R₂₀, R₂₂ and R₂₃ are independently selectedfrom:

-   -   a) (C₁-C₄)alkyl-O— unsubstituted or substituted by 1, 2 or 3        halo;    -   b) (C₁-C₄)alkyl unsubstituted or substituted by OH,        —O—(C₁-C₂)alkyl or 1, 2 or 3 halo,    -   c) HOC(O)—(CH₂)_(n)—,    -   d) H₃C—C(O)(CH₂)_(n)—,    -   e) H₃C—O—C(O)(CH₂)_(n),    -   f) ═O    -   g) R₂₅(R₂₄)N—, wherein R₂₄ is H, (C₁-C₂)alkyl unsubstituted or        substituted by 1, 2 or 3 halo, R₂₅ is H, (C₁-C₂)alkyl        unsubstituted or substituted by 1, 2 or 3 halo,    -   h) OH

wherein n is 0 or 1,

and wherein:

-   -   substituent a) to h) of said heterocyclyl or halo-substituted        heterocyclyl is not present on the ring atoms adjacent to the        ring atom to which the heterocyclyl or halo-substituted        heterocyclyl is joined to the remainder of the molecule, and        preferably, when said heterocyclyl or halo-substituted        heterocyclyl is a 6 membered ring, it has 0 or 1 substituent        selected from a) to h) in the meta or para position, preferably        para, relative to the remainder of the molecule; and    -   said heterocyclyl is linked to the remainder of the compound via        a R₁ ring nitrogen atom, or a R₁ ring carbon atom which is        double bonded to an adjacent ring atom;

or R₁ is heteroaryl, wherein said heteroaryl is a 5 or 6 membered fullyunsaturated monocyclic group comprising ring carbon atoms and 1 or 2ring heteroatoms independently selected from N, O and S, preferably N,wherein the total number of ring S atoms does not exceed 1, and thetotal number of ring O atoms does not exceed 1, wherein said heteroarylis unsubstituted or substituted by 1 or 2 substituents independentlyselected from R₂₁ and R₃₀, wherein R₂₁ and R₃₀ are independentlyselected from (C₁-C₂)alkyl, and said (C₁-C₂)alkyl is unsubstituted orsubstituted by 1, 2 or 3 halo, and wherein preferably, said alkyl orhalo-alkyl substituent is not present on the R₁ ring atoms adjacent tothe R₁ ring atom to which the heteroaryl is joined to the remainder ofthe molecule, and more preferably, when heteroaryl is a 6-membered ring,said alkyl or halo-alkyl substituent is in the ring para positionrelative to the rest of the molecule.

Embodiment 5. A compound of formula (I) or a pharmaceutically acceptablesalt thereof, according to any of Embodiments 1 to 3, wherein R₁ isselected from:

R₃₃ is F;

R₁₅ is halo, azetidinyl or pyrrolidinyl, wherein said azetidinyl andpyrrolidinyl are linked to the rest of the molecule via the N atom, andare unsubstituted or substituted by 1 or 2 F;

R₁₆ is R₂₅(R₂₄)N—, wherein R₂₄ is H or (C₁-C₂)alkyl, R₂₅ is H or(C₁-C₂)alkyl unsubstituted or substituted by 1, 2 or 3 halo, inparticular F;

R₁₇ is halo

R₁₆ is halo;

R₁₉ is halo;

R₂₀ is halo;

R₂₁ is (C₁-C₂)alkyl;

R₂₂ and R₂₃ are each independently selected from:

-   -   (C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo,    -   HOC(O)—(CH₂)_(n)—,    -   H₃C—C(O)(CH₂)_(n)—,    -   (H₃C)₃C—O—C(O)(CH₂)_(n)—;    -   wherein n is 0, 1 or 2;

and

R₃₀ is CH₃.

Embodiment 6. A compound of formula (I) or a pharmaceutically acceptablesalt thereof, according to any of Embodiments 1 to 5, wherein R₁ isselected from:

R₁₅ is F;

R₁₆ is R₂₅(R₂₄)N—;

R₁₇ is F;

R₁₈ is F;

R₁₉ is F;

R₂₀ is F;

R₂₁ is CH₃;

R₂₂ is CF₃, CHF₂CH₂, HOC(O)—CH₂—, H₃C—C(O)—, (H₃C)₃C—O—C(O)—;

R₂₃ is CF₃, CHF₂CH₂—, (H₃C)₃C—O—C(O)—;

R₂₄ is CH₃; and

R₂₅ is CHF₂CH₂—.

Embodiment 7. A compound of formula (I) or a pharmaceutically acceptablesalt thereof, according to any of Embodiments 1 to 6, wherein R₂ is themoiety:

R₆ is selected from:

-   -   H,    -   halo,    -   (C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo,    -   (C₃-C₅)cycloalkyl unsubstituted or substituted by 1, 2 or 3        halo,    -   —O—(C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo,    -   OH, and    -   CN;

R₈ is selected from H, halo, and (C₁-C₄)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo,

R₉ is selected from H, O—CH₃, OH, CN, CH₃ and halo;

R₂₈ is selected from:

-   -   SF₅,    -   H,    -   —C(O)H,    -   halo,    -   (C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo,    -   (C₁-C₄)alkynyl,    -   (C₁-C₄)alkenyl,    -   (C₃-C₅)cycloalkyl unsubstituted or substituted by 1, 2 or 3        halo, and    -   OCF₃;

and X is selected from C—R₇ and N, wherein R₇ is H or halo.

Embodiment 8. A compound of formula (I) or a pharmaceutically acceptablesalt thereof, according to any of Embodiments 1 to 7, wherein R₂ is themoiety:

wherein

R_(e) is selected from H, halo, (C₁-C₄)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo;

R₈ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₉ is selected from H, O—CH₃, OH, CN, CH₃ and halo;

R₂₈ is selected from SF₅, halo, (C₁-C₄)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo and —C(O)H;

X is selected from C—R₇ and N; and

R₇ is selected from H and halo.

Embodiment 9. A compound of formula (I) or a pharmaceutically acceptablesalt thereof, according to any of Embodiments 1 to 8, wherein R₂ is themoiety:

R₆ is selected from H, Cl, CH₃, F and Br;

R₈ is selected from H, Cl, F and CF₃;

R₉ is selected from H, CH₃ and Cl;

R₂₆ is selected from CF₃, CF₂H, —CH₂CH₃, Cl, SF₅, Br and —C(O)H;

X is selected from C—R₇ and N; and

R₇ is selected from H and F.

Embodiment 10. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 9, whereinR₂₆ is H and R₂₇ is H.

Embodiment 11. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 10,wherein R₃ is (C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3substituents independently selected from halo and OH.

Embodiment 12. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 11,wherein R₃ is (C₁-C₂)alkyl unsubstituted or substituted by 1, 2 or 3substituents independently selected from halo and OH, preferably —CH₂CH₃or CH₃, more preferably —CH₂CH₃.

Embodiment 13. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 12,wherein Y is N and Y

is Y linked by a single bond.

Embodiment 14. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 13,wherein K

is K linked by a single bond, and K is selected from —CH₂—, —CH₂CH₂—,—NH— and a bond (to form a 5-membered ring:

J is N, and A is a linker selected from —C(O)—, —S(O)—, —S(O)₂—, and

Embodiment 15. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 14,wherein K

is K linked by a single bond, K is —CH₂—, J is N, and A is a linkerselected from —C(O)—, —S(O)—, —S(O)₂—, and

Embodiment 16. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 15,wherein A is a linker selected from —C(O)— and —S(O)₂—, preferably—C(O)—.

Embodiment 17. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 16,wherein R₅ is independently selected from:

-   -   —(C₁-C₄)alkyl, preferably methyl,    -   and wherein two R₅ substituents on the same ring carbon atom may        join, together with the carbon atom to which they are attached,        to form a (C₃-C₄)cycloalkyl spiro ring or a 3 or 4-membered        heterocyclyl spiro ring, wherein said heterocyclyl spiro ring        contains ring carbon ring atoms and one ring heteroatom selected        from O, N and S,    -   when K        J is a carbon-nitrogen single bond, a R₅ substituent on K and on        the adjacent carbon atom may join to form ring C:

-   -   wherein ring C is a fused (C₃-C₆)cycloalkyl ring, in particular        a fused cyclobutyl ring, a fused (C₃-C₆)heterocyclyl ring or a        fused phenyl ring, wherein said fused (C₃-C₆)heterocyclyl ring        contains ring carbon atoms and one ring heteroatom selected from        O, N and S,    -   and wherein when K is —CH₂— and J is N, two R₅ substituents may        join to form a (C₁-C₃)alkylene bridge or a heteroalkylene        bridge, wherein said heteroalkylene bridge is one heteroatom        selected from N and O, or is —CH₂—O—CH₂—.

Embodiment 18. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 17,wherein R₅ is independently selected from:

-   -   —(C₁-C₄)alkyl, preferably methyl,    -   when K        J is a carbon-nitrogen single bond, a R₅ substituent on K and on        the adjacent carbon atom may join to form ring C:

-   -   wherein ring C is a fused (C₃-C₆)cycloalkyl ring, in particular        a fused cyclobutyl ring, or a fused (C₃-C₆)heterocyclyl ring,        wherein said fused (C₃-C₆)heterocyclyl ring contains ring carbon        atoms and one ring heteroatom selected from O, N and S,    -   and wherein when K is —CH₂— and J is N, two R₅ substituents may        join to form a (C₁-C₃)alkylene bridge or a heteroalkylene        bridge, wherein said heteroalkylene bridge is one heteroatom        selected from N and O, or is —CH₂—O—CH₂—.

Embodiment 19. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 18,wherein R₅ is independently selected from:

-   -   —(C₁-C₂)alkyl, preferably methyl, and    -   when K        J is a carbon-nitrogen single bond, a R₅ substituent on K and on        the adjacent carbon atom may join to form ring C:

-   -   wherein ring C is a fused (C₃-C₄)cycloalkyl ring, in particular        a fused cyclobutyl ring.

Embodiment 20. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 19,wherein R₅ is independently selected from:

-   -   CH₃, and y is 1 or 2, and    -   when k        J is a carbon-nitrogen single bond, a R₅ substituent on K and on        the adjacent carbon atom may join to form ring C:

-   -   wherein ring C is a fused cyclobutyl ring.

Embodiment 21. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 20,wherein y is 0, 1, 2 or 3, preferably 0, 1, or 2.

Embodiment 22. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 21,wherein R₄ is selected from:

wherein

R₁₀ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents, —O—(C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents;

R₁₁ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents;

R₁₂ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents;

R₁₃ is selected from H, —S—CH₃, halo, (C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents; and

R₁₄ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents, O—(C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents, and cyclopropyl.

Embodiment 23. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 22,wherein R₄ is selected from:

wherein

R₁₀ is selected from H, F, Cl, CH₃ and OCF₃;

R₁₁ is selected from H, Cl, F, and CH₃;

R₁₂ is selected from H, Cl and CH₃;

R₁₃ is selected from H, —S—CH₃ and CH₃; and

R₁₄ is selected from H, CH₃, —CH₂CH₃, cyclopropyl, —OCHF₂, OCF₃ and Cl.

Embodiment 24. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 23,wherein formula (I) is formula 1a:

(Preferably, formula (I) is formula 1a)

Embodiment 25. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 23,wherein formula (I) is formula 1b:

Embodiment 26. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 23,wherein formula (I) is formula 1c:

Embodiment 27. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 23,wherein formula (I) is formula 1d:

Embodiment 28. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 23,wherein formula (I) is formula 1e:

Embodiment 29. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 23,wherein formula (I) is formula 1f:

Embodiment 30. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 24,wherein formula (I) is formula 1g:

More preferably, formula (I) is formula 1g.

Embodiment 31. A compound of formula (I) or a pharmaceuticallyacceptable salt thereof, according to any of Embodiments 1 to 24, or 30,wherein formula (I) is formula 1h:

Most preferably, formula (I) is formula 1h.

Embodiment 32. A compound of formula (Ig) or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1, 24 or 30,

wherein R₁ is selected from:

R₁₅ is H or F;

R₁₆ is H or R₂₅(R₂₄)N—;

R₁₇ is H or F;

R₁₈ is H or F;

R₁₉ is H or F;

R₂₀ is H or F;

R₂₁ is H or CH₃;

R₂₂ is H, CF₃, CHF₂CH₂, HOC(O)—CH₂—, H₃C—C(O)—, (H₃C)₃C—O—C(O)—;

R₂₃ is H, CF3, CHF₂CH₂—, (H₃C)₃C—O—C(O)—;

R₂₄ is CH₃;

R₂₅ is CHF₂CH₂—;

R₂₆ is CH₃, H or deuterium;

R₂₇ is H or deuterium;

R₂ is the moiety:

R₆ is selected from H, Cl, CH₃, F and Br;

R₈ is selected from H, Cl, F and CF₃;

R₉ is selected from H, CH₃ and Cl;

R₂₈ is selected from CF₃, CF₂H, —CH₂CH₃, Cl, SF₅, Br and —C(O)H;

X is selected from C—R₇ and N;

R₇ is selected from H and F;

R₃ is selected from CH₃, CH₂CH₃, cyclopropyl, hydroxyethyl;

R₄ is selected from:

wherein

R₁₀ is selected from H, F, Cl, CH₃ and OCF₃;

R₁₁ is selected from H, Cl, F, and CH₃;

R₁₂ is selected from H, Cl and CH₃;

R₁₃ is selected from H, —S—CH₃ and CH₃;

R₁₄ is selected from H, CH₃, —CH₂CH₃, cyclopropyl, —OCHF₂, OCF₃ and Cl;

Y is N or CH, preferably N,

y is 0, 1 or 2;

R₅ is CH₃, or alternatively, two R₅ groups on adjacent carbon atomsjoin, along with the carbon atoms to which they are attached, to form afused cyclobutyl ring:

and optionally, wherein one or more H atoms of the ring:

are replaced by deuterium;

* indicates a point of attachment.

Embodiment 33. A compound of formula (I), according to embodiment 1, 24or 30, or a pharmaceutically acceptable salt thereof:

wherein

R₁ is selected from:

R₁₅ is H or F;

R₁₆ is H or R₂₅(R₂₄)N—;

R₁₇ is H or F;

R₁₈ is H or F;

R₁₉ is H or F;

R₂₀ is H or F;

R₂₁ is H or CH₃;

R₂₂ is H, CF₃, CHF₂CH₂, HOC(O)—CH₂—, H₃C—C(O)—, (H₃C)₃C—O—C(O)—;

R₂₃ is H, CF₃, CHF₂CH₂—, (H₃C)₃C—O—C(O)—;

R₂₄ is CH₃;

R₂₅ is CHF₂CH₂—;

R₂₆ is CH₃, H or deuterium;

R₂₇ is H or deuterium;

R₂ is the moiety:

wherein

R₆ is selected from H, Cl, CH₃, F, and Br;

R₆ is selected from H, Cl, F and CF₃;

R₉ is selected from H, CH₃ and Cl;

R₂₆ is selected from CF₃, CF₂H, —CH₂CH₃, Cl, SF₅, Br and —C(O)H;

X is selected from C—R₇ and N;

R₇ is selected from H and F;

R₃ is selected from CH₃, CH₂CH₃, cyclopropyl and hydroxyethyl;

R₄ is selected from:

wherein

R₁₀ is selected from H, F, Cl, CH₃ and OCF₃;

R₁₁ is selected from H, Cl, F, and CH₃;

R₁₂ is selected from H, Cl and CH₃;

R₁₃ is selected from H and CH₃;

R₁₄ is selected from H, CH₃, —CH₂CH₃, cyclopropyl, —OCHF₂, OCF₃ and Cl;

y is 0, 1 or 2;

R₅ is CH₃; or alternatively, two R₅ groups on adjacent carbon atomsjoin, along with the carbon atoms to which they are attached, to form afused cyclobutyl ring:

and

* indicates a point of attachment.

Embodiment 34. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1 to 33,wherein R₁ is selected from:

Embodiment 35. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 34, wherein R₁ isselected from:

Embodiment 36. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 35, wherein R₁ isselected from:

Embodiment 37. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 36, wherein R₁ is:

Embodiment 38. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-37, whereinR₂₈ is selected from CF₃, SF₅ and Br.

Embodiment 39. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 38, wherein R₂₈ isselected from CF₃.

Embodiment 40. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-39, wherein Xis CR₇.

Embodiment 41. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-40, whereinR₇ is H.

Embodiment 42. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-41, whereinR₆ is Cl or CH₃.

Embodiment 43. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 42, wherein Re is Cl.

Embodiment 44. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-43, whereinR₈ is F or H.

Embodiment 45. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 44, wherein R₈ is H.

Embodiment 46. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-45, whereinR₉ is H.

Embodiment 47. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-46, whereinR₂₆ is H.

Embodiment 48 A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-47, whereinR₂₇ is H.

Embodiment 49. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 1, wherein R₂₆ and R₂₇are both deuterium.

Embodiment 50. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-49, whereinR₃ is —CH₂—CH₃.

Embodiment 51. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-50, wherein yis 0.

Embodiment 52. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-51, whereinthe moiety:

is selected from

Embodiment 53. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 52, wherein the moiety:

is selected from:

or is selected from:

Embodiment 54. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-51, wherein themoiety:

is selected from:

Embodiment 55. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-54, whereinthe moiety:

is selected from:

Embodiment 56. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 55, wherein the moiety:

is selected from:

Embodiment 57. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 55, wherein the moiety:

is selected from:

Embodiment 58. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 56, wherein the moiety:

is selected from:

Embodiment 59. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-58, whereinR₁₀ is H, F or Cl.

Embodiment 60. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-59, whereinR₁₁ is H or CH₃.

Embodiment 61. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-60, whereinR₁₂ is H.

Embodiment 62. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-61, whereinR₁₃ is H.

Embodiment 63. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-62, whereinR₁₄ is CH₃ or H.

Embodiment 64. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-63, whereinR₁₄ is CH₃.

Embodiment 65. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-64, whereinR₄ is selected from:

Embodiment 66. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 65, wherein R₄ isselected from:

Embodiment 67. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 66, wherein R₄ isselected from:

Embodiment 68. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1 and 24 to 31,wherein R₁ is selected from:

R₁₅ is H or F;

R₁₆ is H or R₂₅(R₂₄)N—;

R₁₇ is H or F;

R₁₈ is H or F;

R₁₉ is H or F;

R₂₀ is H or F;

R₂₁ is H or CH₃;

R₂₂ is H, CF₃, CHF₂CH₂, HOC(O)—CH₂—, H₃C—C(O)—, (H₃C)₃C—O—C(O)—;

R₂₃ is H, CF3, CHF₂CH₂—, (H₃C)₃C—O—C(O)—;

R₂₄ is CH₃;

R₂₅ is CHF₂CH₂—; and

R₄ is selected from:

wherein

R₁₀ is selected from H, F, Cl, CH₃ and OCF₃;

R₁₁ is selected from H, Cl, F, and CH₃;

R₁₂ is selected from H, Cl and CH₃;

R₁₃ is selected from H and CH₃;

R₁₄ is selected from H, CH₃, —CH₂CH₃, cyclopropyl, —OCHF₂, OCF₃ and Cl;

or a pharmaceutically acceptable salt thereof.

Embodiment 69. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1 and 24 to 31,wherein R₁ is selected from:

and the moiety:

is selected from:

in particular wherein

the moiety:

is selected from:

and

R₄ is selected from:

Embodiment 70. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1 and 24 to 31,wherein R₁ is selected from:

the moiety:

is selected from:

the moiety:

is selected from:

in particular

and R₄ is selected from:

in particular:

Embodiment 71. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1, 24, 30 and31, wherein the compound is selected from:

Embodiment 72. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1 or 39, wherein thecompound is selected from:

Embodiment 73. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1, 39 or 40, whereinthe compound is selected from:

Embodiment 74. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-73, whereinthe compound is in non-zwitterionic form.

Embodiment 75. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-73, whereinthe compound is in zwitterionic form.

Embodiment 76. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-73, whereinthe compound is a mixture of zwitterionic and non-zwitterionic forms.

Embodiment 77. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-73, whereinthe R₄ group is present in non-zwitterionic form (d) or (e):

Embodiment 78. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-73, whereinR₄ is present in a zwitterionic form selected from:

Embodiment 79. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-73, whereinR₄ is present as a mixture of zwitterionic forms (a) and (b) accordingto embodiment 78.

Embodiment 80. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-73, whereinR₄ is present as a mixture of:

-   -   Non-zwitterionic form (e) and zwitterionic forms (a) or (b),

or

-   -   Non-zwitterionic form (e) and zwitterionic forms (a) and (b)

Embodiment 81. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-73, whereinR₄ is in zwitterionic form (c):

Embodiment 82. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-73, whereinR₄ is present as a mixture of both zwitterionic form (c) andnon-zwitterionic form (d):

Embodiment 83. A compound of formula (I), according to any ofembodiments 1, 24, 30 or 31, wherein the compoundN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideis:

in non-zwitterionic form:

or zwitterionic form:

or zwitterionic form:

or a mixture of any two or three of said forms.

Embodiment 84. A compound of formula (I), according to any ofembodiments 1, 24, 30 or 31, wherein the compound(R)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideis:

in non-zwitterionic form:

or in zwitterionic form:

or in zwitterionic form:

or a mixture of any two or three of said forms.

Embodiment 85. A compound of formula (I), according to any ofembodiments 1, 24, 30 or 31, wherein the compoundN-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideis:

in non-zwitterionic form:

or in zwitterionic form:

or in zwitterionic form:

or a mixture of any two or three of said forms.

Embodiment 86. A compound of formula (I), according to any ofembodiments 1, 24, 30 or 31, wherein the compound2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamideis:

in non-zwitterionic form:

or in zwitterionic form:

or in zwitterionic form:

or a mixture of any two or three of said forms.

Embodiment 87: A compound of formula (I), according to any ofembodiments 1, 24, 30 or 31, wherein the compoundN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideis:

in non-zwitterionic form:

or in zwitterionic form:

or a mixture of said forms.

Embodiment 88. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-73, whereinthe compound is a sodium salt.

Embodiment 89. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-88, whereinthe compound is in amorphous form. For example, the compound is thesodium salt in amorphous form.

Embodiment 90. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to any of embodiments 1-88, incrystalline form.

Embodiment 91. A compound of formula (I) according to any of embodiments1, 24, 30 and 31, wherein the compound is:

in crystalline form.

Embodiment 92. A compound of formula (I) according to any of embodiments1, 24, 30 and 31, wherein the compound is:

in crystalline form.

Embodiment 93. A compound of formula (I) according to any of embodiments1, 24, 30 and 31, wherein the compound is:

in crystalline form.

Embodiment 94. A compound of formula (I) according to any of embodiments1, 24, 30 and 31, wherein the compound is:

in crystalline form.

Embodiment 95. A compound of formula (I) according to any of embodiments1, 24, 30 and 31, wherein the compound is:

in crystalline form.

Embodiment 96. A compound of formula (I) according to embodiments 91-95,wherein the compound is in substantially pure form.

Embodiment 97. A compound of formula (I) according to embodiment 1,wherein the crystalline form according to embodiment 91 is characterizedby a X-ray powder diffraction pattern comprising 4 or more 2θ valuesselected from the group consisting of 11.85±0.2, 13.71±0.2, 14.46±0.2,15.33±0.2, 17.03±0.2, 18.33±0.2, 19.98±0.2, 22.42±0.2, 22.95±0.2 and27.20±0.2, at a temperature of about 22° C., or comprising 4 or more 2θvalues selected from the group consisting of 6.78±0.2, 8.97±0.2,11.88±0.2, 13.55±0.2, 13.74±0.2, 14.48±0.2, 15.34±0.2, 16.83±0.2,17.03±0.2, 18.30±0.2, 19.49±0.2, 19.94±0.2, 21.28±0.2, 21.51±0.2,22.38±0.2, 22.91±0.2, 23.27±0.2, 25.41±0.2, 27.26±0.2, 29.03±0.2,29.78±0.2 and 29.96±0.2, at a temperature of about 22° C.

Embodiment 98. A compound of formula (I) according to embodiment 1,wherein the crystalline form according to embodiment 91 is furthercharacterized by a X-ray powder diffraction pattern comprising 5 or more2θ values selected from the group consisting of 11.85±0.2, 13.71±0.2,14.46±0.2, 15.33±0.2, 17.03±0.2, 18.33±0.2, 19.98±0.2, 22.42±0.2,22.95±0.2 and 27.20±0.2, at a temperature of about 22° C., or comprising5 or more 20 values selected from the group consisting of 6.78±0.2,8.97±0.2, 11.88±0.2, 13.55±0.2, 13.74±0.2, 14.48±0.2, 15.34±0.2,16.83±0.2, 17.03±0.2, 18.30±0.2, 19.49±0.2, 19.94±0.2, 21.28±0.2,21.51±0.2, 22.38±0.2, 22.91±0.2, 23.27±0.2, 25.41±0.2, 27.26±0.2,29.03±0.2, 29.78±0.2 and 29.96±0.2, at a temperature of about 22° C.

Embodiment 99. A compound of formula (I) according to embodiment 1,wherein the crystalline form according to embodiment 91 is characterizedby a X-ray diffraction pattern substantially the same as the X-raypowder diffraction pattern shown in FIG. 1 or FIG. 7 .

Embodiment 100. A compound of formula (I) according to embodiment 1,wherein the crystalline form according to embodiment 92 is characterizedby a X-ray powder diffraction pattern comprising 4 or more 2θ valuesselected from the group consisting of 10.29±0.2, 13.31±0.2, 14.01±0.2,15.26±0.2 and 17.34±0.2 at a temperature of about 22° C.

Embodiment 101. A compound of formula (I) according to embodiment 1,wherein the crystalline form according to embodiment 92 is furthercharacterized by a X-ray powder diffraction pattern comprising 5 2θvalues, selected from the group consisting of 10.29±0.2, 13.31±0.2,14.01±0.2, 15.26±0.2 and 17.34±0.2 at a temperature of about 22° C.

Embodiment 102. A compound of formula (I) according to embodiment 1,wherein the crystalline form according to embodiment 92 is characterizedby a X-ray diffraction pattern substantially the same as the X-raypowder diffraction spectrum shown in FIG. 2 .

Embodiment 103. A compound of formula (I) according to embodiment 1,wherein the crystalline form according to embodiment 93 is characterizedby a X-ray powder diffraction pattern comprising 4 or more 2θ valuesselected from the group consisting of 9.45±0.2, 12.75±0.2, 13.28±0.2,21.69±0.2, 25.25±0.2 and 26.85±0.2 at a temperature of about 22° C.

Embodiment 104. A compound of formula (I) according to embodiment 1,wherein the crystalline form according to embodiment 93 is furthercharacterized by a X-ray powder diffraction pattern comprising 5 or more2θ values selected from the group consisting of 9.45±0.2, 12.75±0.2,13.28±0.2, 21.69±0.2, 25.25±0.2 and 26.85±0.2 at a temperature of about22° C.

Embodiment 105. A compound of formula (I) according to embodiment 1,wherein the crystalline form according to embodiment 93 is characterizedby a X-ray diffraction pattern substantially the same as the X-raypowder diffraction spectrum shown in FIG. 3 .

Embodiment 106. A compound of formula (I) according to embodiment 1,wherein the crystalline form according to embodiment 94 is characterizedby a X-ray powder diffraction pattern comprising 4 or more 2θ valuesselected from the group consisting of 11.81±0.2, 13.75±0.2, 14.45±0.2,15.32±0.2, 17.04±0.2, 17.40±0.2, 18.27±0.2, 19.95±0.2, 22.92±0.2 and27.13±0.2 at a temperature of about 22° C.

Embodiment 107. A compound of formula (I) according to embodiment 1,wherein the crystalline form according to embodiment 94 is furthercharacterized by a X-ray powder diffraction pattern comprising 5 or more2θ values selected from the group consisting of 11.81±0.2, 13.75±0.2,14.45±0.2, 15.32±0.2, 17.04±0.2, 17.40±0.2, 18.27±0.2, 19.95±0.2,22.92±0.2 and 27.13±0.2 at a temperature of about 22° C.

Embodiment 108. A compound of formula (I) according to embodiment 1,wherein the crystalline form according to embodiment 94 is characterizedby a X-ray diffraction pattern substantially the same as the X-raypowder diffraction spectrum shown in FIG. 4 .

Embodiment 109. A compound of formula (I) according to embodiment 1,wherein the crystalline form according to embodiment 95 is characterizedby a X-ray powder diffraction pattern comprising 4 or more 2θ valuesselected from the group consisting of 13.20±0.2, 14.78±0.2, 15.97±0.2,16.91±0.2, 19.95±0.2, 20.85±0.2, 24.43±0.2, 25.47±0.2 and 31.06±0.2 at atemperature of about 22° C.

Embodiment 110. A compound of formula (I) according to embodiment 1,wherein the crystalline form according to embodiment 95 is furthercharacterized by a X-ray powder diffraction pattern comprising 5 or more2θ values selected from the group consisting of 13.20±0.2, 14.78±0.2,15.97±0.2, 16.91±0.2, 19.95±0.2, 20.85±0.2, 24.43±0.2, 25.47±0.2 and31.06±0.2 at a temperature of about 22° C.

Embodiment 111. A compound of formula (I) according to embodiment 1,wherein the crystalline form according to embodiment 95 is characterizedby a X-ray diffraction pattern substantially the same as the X-raypowder diffraction spectrum shown in FIG. 5 .

Embodiment 112. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 1, wherein formula (I)is formula 1a:

wherein R₁ is selected from:

R₃₃ is F;

R₁₅ is halo, azetidinyl or pyrrolidinyl, wherein said azetidinyl andpyrrolidinyl are linked to the rest of the molecule via the N atom, andare unsubstituted or substituted by 1 or 2 F;

R₁₆ is R₂₅(R₂₄)N—, wherein R₂₄ is H or (C₁-C₂)alkyl, R₂₅ is H or(C₁-C₂)alkyl;

R₁₇ is halo

R₁₈ is halo;

R₁₉ is halo;

R₂₀ is halo;

R₂₁ is (C₁-C₂)alkyl;

R₂₂ and R₂₃ are each independently selected from:

-   -   (C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo,    -   HOC(O)—(CH₂)_(n)—,    -   H₃C—C(O)(CH₂)_(n)—,    -   (H₃C)₃C—O—C(O)(CH₂)_(n)—;    -   wherein n is 0, 1 or 2;

and

R₃₀ is CH₃.

R₂ is the moiety:

wherein

R₆ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₈ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₉ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₂₈ is selected from SF₅, halo, (C₁-C₄)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo and —C(O)H;

X is selected from C—R₇ and N; and

R₇ is selected from H and halo;

R₂₆ is H and R₂₇ is H;

R₃ is —CH₂CH₃ or CH₃;

A is a linker selected from —C(O)— and —S(O)₂—, preferably —C(O)—;

and

R₄ is selected from:

wherein

R₁₀ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents, —O—(C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents;

R₁₁ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents;

R₁₂ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents;

R₁₃ is selected from H, —S—CH₃ halo, (C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents; and

R₁₄ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents, O—(C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents, cyclopropyl.

Preferably, Formula (I), or 1a, is 1g:

and more preferably 1h:

Embodiment 113. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 1, wherein formula (I)is formula 1b:

wherein R₁ is selected from:

R₃₃ is F;

R₁₅ is halo, azetidinyl or pyrrolidinyl, wherein said azetidinyl andpyrrolidinyl are linked to the rest of the molecule via the N atom, andare unsubstituted or substituted by 1 or 2 F;

R₁₆ is R₂₅(R₂₄)N—, wherein R₂₄ is H or (C₁-C₂)alkyl, R₂₅ is H or(C₁-C₂)alkyl;

R₁₇ is halo

R₁₈ is halo;

R₁₉ is halo;

R₂₀ is halo;

R₂₁ is (C₁-C₂)alkyl;

R₂₂ and R₂₃ are each independently selected from:

-   -   (C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo,    -   HOC(O)—(CH₂)_(n)—,    -   H₃C—C(O)(CH₂)_(n)—,    -   (H₃C)₃C—O—C(O)(CH₂)_(n)—;    -   wherein n is 0, 1 or 2;

and

R₃₀ is CH₃.

R₂ is the moiety:

wherein

R₆ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₈ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₉ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₂₈ is selected from SF₅, halo, (C₁-C₄)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo and —C(O)H;

X is selected from C—R₇ and N; and

R₇ is selected from H and halo;

R₂₆ is H and R₂₇ is H;

R₃ is —CH₂CH₃ or CH₃;

A is a linker selected from —C(O)— and —S(O)₂—, preferably —C(O)—;

and

R₄ is selected from:

wherein

R₁₀ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents, —O—(C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents;

R₁₁ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents;

R₁₂ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents;

R₁₃ is selected from H, —S—CH₃ halo, (C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents; and

R₁₄ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents, O—(C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents, cyclopropyl.

Formula 1b is preferably 1b1:

Particularly, where Y is N.

Embodiment 114. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 1, wherein formula (I)is formula 1c:

wherein R₁ is selected from:

R₃₃ is F;

R₁₅ is halo, azetidinyl or pyrrolidinyl, wherein said azetidinyl andpyrrolidinyl are linked to the rest of the molecule via the N atom, andare unsubstituted or substituted by 1 or 2 F;

R₁₆ is R₂₅(R₂₄)N—, wherein R₂₄ is H or (C₁-C₂)alkyl, R₂₅ is H or(C₁-C₂)alkyl;

R₁₇ is halo

R₁₈ is halo;

R₁₉ is halo;

R₂₀ is halo;

R₂₁ is (C₁-C₂)alkyl;

R₂₂ and R₂₃ are each independently selected from:

-   -   (C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo,    -   HOC(O)—(CH₂)_(n)—,    -   H₃C—C(O)(CH₂)_(n)—,    -   (H₃C)₃C—O—C(O)(CH₂)_(n)—;    -   wherein n is 0, 1 or 2;

and

R₃₀ is CH₃.

R₂ is the moiety:

wherein

R₆ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₈ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₉ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₂₆ is selected from SF₅, halo, (C₁-C₄)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo and —C(O)H;

X is selected from C—R₇ and N; and

R₇ is selected from H and halo;

R₂₆ is H and R₂₇ is H;

R₃ is —CH₂CH₃ or CH₃;

A is a linker selected from —C(O)— and —S(O)₂—, preferably —C(O)—;

and

R₄ is selected from:

wherein

R₁₀ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents, —O—(C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents;

R₁₁ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents;

R₁₂ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents;

R₁₃ is selected from H, —S—CH₃ halo, (C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents; and

R₁₄ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents, O—(C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents, cyclopropyl.

Formula 1c, is preferably 1c1:

particularly where Y is N.

Embodiment 115. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 1, wherein formula (I)is formula 1d:

wherein R₁ is selected from:

R₃₃ is F;

R₁₅ is halo, azetidinyl or pyrrolidinyl, wherein said azetidinyl andpyrrolidinyl are linked to the rest of the molecule via the N atom, andare unsubstituted or substituted by 1 or 2 F;

R₁₆ is R₂₅(R₂₄)N—, wherein R₂₄ is H or (C₁-C₂)alkyl, R₂₅ is H or(C₁-C₂)alkyl;

R₁₇ is halo

R₁₈ is halo;

R₁₉ is halo;

R₂₀ is halo;

R₂₁ is (C₁-C₂)alkyl;

R₂₂ and R₂₃ are each independently selected from:

-   -   (C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo,    -   HOC(O)—(CH₂)_(n)—,    -   H₃C—C(O)(CH₂)_(n)—,    -   (H₃C)₃C—O—C(O)(CH₂)_(n)—;    -   wherein n is 0, 1 or 2;

and

R₃₀ is CH₃.

R₂ is the moiety:

wherein

R₆ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₈ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₉ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₂₈ is selected from SF₅, halo, (C₁-C₄)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo and —C(O)H;

X is selected from C—R₇ and N; and

R₇ is selected from H and halo;

R₂₆ is H and R₂₇ is H;

R₃ is —CH₂CH₃ or CH₃;

A is a linker selected from —C(O)— and —S(O)₂—, preferably —C(O)—;

and

R₄ is selected from:

wherein

R₁₀ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents, —O—(C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents;

R₁₁ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents;

R₁₂ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents;

R₁₃ is selected from H, —S—CH₃ halo, (C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents; and

R₁₄ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents, O—(C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents, cyclopropyl.

Formula 1d, is preferably 1d1:

Particularly, where Y is N.

Embodiment 116. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 1, wherein formula (I)is formula 1e:

wherein R₁ is selected from:

R₃₃ is F;

R₁₅ is halo, azetidinyl or pyrrolidinyl, wherein said azetidinyl andpyrrolidinyl are linked to the rest of the molecule via the N atom, andare unsubstituted or substituted by 1 or 2 F;

R₁₆ is R₂₅(R₂₄)N—, wherein R₂₄ is H or (C₁-C₂)alkyl, R₂₅ is H or(C₁-C₂)alkyl;

R₁₇ is halo

R₁₈ is halo;

R₁₉ is halo;

R₂₀ is halo;

R₂₁ is (C₁-C₂)alkyl;

R₂₂ and R₂₃ are each independently selected from:

-   -   (C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo,    -   HOC(O)—(CH₂)_(n)—,    -   H₃C—C(O)(CH₂)_(n)—,    -   (H₃C)₃C—O—C(O)(CH₂)_(n)—;    -   wherein n is 0, 1 or 2;

and

R₃₀ is CH₃.

R₂ is the moiety:

wherein

R₆ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₈ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₉ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₂₈ is selected from SF₅, halo, (C₁-C₄)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo

and —C(O)H;

X is selected from C—R₇ and N; and

R₇ is selected from H and halo;

R₂₆ is H and R₂₇ is H;

R₃ is —CH₂CH₃ or CH₃;

A is a linker selected from —C(O)— and —S(O)₂—, preferably —C(O)—;

and

R₄ is selected from:

wherein

R₁₀ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents, —O—(C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents;

R₁₁ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents;

R₁₂ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents;

R₁₃ is selected from H, —S—CH₃ halo, (C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents; and

R₁₄ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents, O—(C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents, cyclopropyl.

Formula 1e, is preferably 1e1:

Particularly, where Y is N.

Embodiment 117. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 1, wherein formula (I)is formula 1f:

wherein R₁ is selected from:

R₃₃ is F;

R₁₅ is halo, azetidinyl or pyrrolidinyl, wherein said azetidinyl andpyrrolidinyl are linked to the rest of the molecule via the N atom, andare unsubstituted or substituted by 1 or 2 F;

R₁₆ is R₂₅(R₂₄)N—, wherein R₂₄ is H or (C₁-C₂)alkyl, R₂₅ is H or(C₁-C₂)alkyl;

R₁₇ is halo

R₁₈ is halo;

R₁₉ is halo;

R₂₀ is halo;

R₂₁ is (C₁-C₂)alkyl;

R₂₂ and R₂₃ are each independently selected from:

-   -   (C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo,    -   HOC(O)—(CH₂)_(n)—,    -   H₃C—C(O)(CH₂)_(n)—,    -   (H₃C)₃C—O—C(O)(CH₂)_(n)—;    -   wherein n is 0, 1 or 2;

and

R₃₀ is CH₃.

R₂ is the moiety:

wherein

R₆ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₈ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₉ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₂₈ is selected from SF₅, halo, (C₁-C₄)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo and —C(O)H;

X is selected from C—R₇ and N; and

R₇ is selected from H and halo;

R₂₆ is H and R₂₇ is H;

R₃ is —CH₂CH₃ or CH₃;

A is a linker selected from —C(O)— and —S(O)₂—, preferably —C(O)—;

and

R₄ is selected from:

wherein

R₁₀ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents, —O—(C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents;

R₁₁ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents;

R₁₂ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents;

R₁₃ is selected from H, —S—CH₃ halo, (C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents; and

R₁₄ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents, O—(C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents, cyclopropyl.

Formula 1f, is preferably 1f1:

Particularly, where Y is N.

Embodiment 1.1. A compound of formula (I) according to embodiment 1, ora pharmaceutically acceptable salt thereof:

wherein (I) is 1 g, and

R₁ is selected from:

R₁₅ is H or F;

R₁₆ is H or R₂₅(R₂₄)N—;

R₁₇ is H or F;

R₁₈ is H or F;

R₁₉ is H or F;

R₂₀ is H or F;

R₂₁ is H or CH₃;

R₂₂ is H, CF₃, CHF₂CH₂, HOC(O)—CH₂—, H₃C—C(O)—, (H₃C)₃C—O—C(O)—;

R₂₃ is H, CF₃, CHF₂CH₂—, (H₃C)₃C—O—C(O)—;

R₂₄ is CH₃;

R₂₅ is CHF₂CH₂—;

R₂₆ is CH₃, H or deuterium;

R₂₇ is H or deuterium;

R₂ is the moiety:

wherein R₆ is selected from H, Cl, CH₃, F, and Br;

R₈ is selected from H, Cl, F and CF₃;

R₉ is selected from H, CH₃ and Cl;

R₂₈ is selected from CF₃, CF₂H, —CH₂CH₃, Cl, SF₅, Br and —C(O)H;

X is selected from C—R₇ and N;

R₇ is selected from H and F;

R₃ is selected from CH₃, CH₂CH₃, cyclopropyl and hydroxyethyl;

R₄ is selected from:

wherein

R₁₀ is selected from H, F, Cl, CH₃ and OCF₃;

R₁₁ is selected from H, Cl, F, and CH₃;

R₁₂ is selected from H, Cl and CH₃;

R₁₃ is selected from H and CH₃;

R₁₄ is selected from H, CH₃, —CH₂CH₃, cyclopropyl, —OCHF₂, OCF₃ and Cl;

y is 0, 1 or 2;

R₅ is CH₃; or alternatively, two R₅ groups on adjacent carbon atomsjoin, along with the carbon atoms to which they are attached, to form afused cyclobutyl ring:

and

* indicates a point of attachment;

or a pharmaceutically acceptable salt thereof.

Embodiment 2.1. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 1, wherein R₁ isselected from:

Embodiment 3.1. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 1 or 2, wherein R₁ isselected from:

Embodiment 4.1. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-3, wherein R₁ isselected from:

Embodiment 5.1. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-4, wherein R₁ is:

Embodiment 6.1. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-5, wherein R²⁸ isselected from CF₃, SF₅ and Br.

Embodiment 7.1. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-6, wherein R²⁸ isselected from CF₃.

Embodiment 8.1. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-7, wherein X is CR₇.

Embodiment 9.1. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-8, wherein R₇ is H.

Embodiment 10.1. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-9, wherein R₆ is Clor CH₃.

Embodiment 11.1. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-10, wherein Re isCl.

Embodiment 12.1. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-11, wherein R₈ is For H.

Embodiment 13.1. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-12, wherein R₈ is H.

Embodiment 14.1. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-13, wherein R₉ is H.

Embodiment 15.1. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-14, wherein R₂₆ isH.

Embodiment 16.1. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-15, wherein R₂₇ isH.

Embodiment 17.1. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-14, wherein R₂₆ andR₂₇ are both deuterium.

Embodiment 18.1. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-18, wherein R₃ is—CH₂—CH₃.

Embodiment 19.1. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-18, wherein y is 0.

Embodiment 20.1. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-18, wherein themoiety:

is selected from

Embodiment 21.1 A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-18 and 20, whereinthe moiety:

is selected from:

Embodiment 22.1 A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-18 and 20, whereinthe moiety:

is selected from:

Embodiment 23.1 A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-22, wherein themoiety:

is selected from:

Embodiment 24.1 A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-23, wherein themoiety:

is selected from:

Embodiment 25.1 A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-24, wherein themoiety:

is selected from:

Embodiment 26.1 A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-25, wherein themoiety:

is selected from:

Embodiment 27.1 A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-26, wherein R₁₀ isH, F or Cl.

Embodiment 28.1 A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-27, wherein R₁₁ is Hor CH₃.

Embodiment 29.1 A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-28, wherein R₁₂ isH.

Embodiment 30.1 A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-29, wherein R₁₃ isH.

Embodiment 31.1 A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-30, wherein R₁₄ isCH₃ or H.

Embodiment 32.1 A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-31, wherein R₁₄ isCH₃.

Embodiment 33.1 A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-32, wherein R₄ isselected from:

Embodiment 34.1 A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-33, wherein R₄ isselected from:

Embodiment 35.1 A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1-34, wherein R₄ isselected from:

Embodiment 36.1 A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiments 1 and 6-26, wherein R₁is selected from:

R₁₅ is H or F;

R₁₆ is H or R₂₅(R₂₄)N—;

R₁₇ is H or F;

R₁₈ is H or F;

R₁₉ is H or F;

R₂₀ is H or F;

R₂₁ is H or CH₃;

R₂₂ is H, CF₃, CHF₂CH₂, HOC(O)—CH₂—, H₃C—C(O)—, (H₃C)₃C—O—C(O)—;

R₂₃ is H, CF3, CHF₂CH₂—, (H₃C)₃C—O—C(O)—;

R₂₄ is CH₃;

R₂₅ is CHF₂CH₂—; and

R₄ is selected from:

wherein

R₁₀ is selected from H, F, Cl, CH₃ and OCF₃;

R₁₁ is selected from H, Cl, F, and CH₃;

R₁₂ is selected from H, Cl and CH₃;

R₁₃ is selected from H and CH₃;

R₁₄ is selected from H, CH₃, —CH₂CH₃, cyclopropyl, —OCHF₂, OCF₃ and Cl;or a pharmaceutically acceptable salt thereof.

Embodiment 37.1 A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 1, wherein R₁ isselected from:

and the moiety:

is selected from:

the moiety:

is selected from:

and

R₄ is selected from:

Embodiment 38.1 A compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to embodiment 37, wherein R₁ isselected from:

the moiety:

is selected from:

the moiety:

is selected from:

in particular

and R₄ is selected from:

in particular:

In a further aspect, the invention is as claimed herein.

In one embodiment, there is provided a compound of formula (I) or apharmaceutically acceptable salt thereof:

wherein

R₁ is:

cycloalkenyl, wherein said cycloalkenyl is a partially unsaturatedmonocyclic ring containing 5 or 6 ring carbon atoms, and saidcycloalkenyl is unsubstituted or substituted by 1, 2, 3 or 4, preferably1 or 2, R₃₃, wherein R₃₃ is halo, and wherein said cycloalkenyl orhalo-substituted cycloalkenyl is substituted by 0, 1 or 2 R₁₅substituents,

or R₁ is heterocyclyl, wherein said heterocyclyl is a 5 or 6 memberedfully saturated or partially unsaturated group comprising ring carbonatoms and 1 or 2 ring heteroatoms independently selected from N, O andS, and wherein said heterocyclyl is unbridged or bridged, and saidbridge is 1 or 2 carbon atoms, wherein said heterocyclyl isunsubstituted or substituted by 1, 2, 3 or 4, preferably 1 or 2, R₃₃,wherein R₃₃ is halo, and wherein said heterocyclyl or halo-substitutedheterocyclyl is substituted by 0, 1 or 2 substituents independentlyselected from R₁₅, R₁₆, R₁₇, R₁₈, R₁₉, R₂₀, R₂₂ and R₂₃,

or R₁ is heteroaryl, wherein said heteroaryl is a 5 or 6 membered fullyunsaturated monocyclic group comprising ring carbon atoms and 1, 2, 3 or4 ring heteroatoms independently selected from N, O and S, preferably 1or 2 ring heteroatoms, wherein the total number of ring S atoms does notexceed 1, and the total number of ring O atoms does not exceed 1,wherein said heteroaryl is unsubstituted or substituted by 1, 2 or 3substituents independently selected from R₂₁ and R₃₀, wherein R₂₁ andR₃₀ are independently selected from halo and (C₁-C₄)alkyl, wherein said(C₁-C₄)alkyl is unsubstituted or substituted by 1, 2 or 3 halo,

and each R₁₅, R₁₆, R₁₇, R₁₈, R₁₉, R₂₀, R₂₂ and R₂₃ is independentlyselected from:

-   -   halo    -   (C₁-C₄)alkyl-O— unsubstituted or substituted by 1, 2 or 3 halo;    -   (C₁-C₄)alkyl unsubstituted or substituted by OH, —O—(C₁-C₂)alkyl        or 1, 2 or 3 halo,    -   HOC(O)—(CH₂)_(n)—,    -   H₃C—C(O)(CH₂)_(n)—,    -   (C₁-C₄)alkyl-O—C(O)(CH₂)_(n),    -   ═O    -   azetidinyl or pyrrolidinyl, wherein said azetidinyl and        pyrrolidinyl are linked to the rest of the molecule via the N        atom, and are each unsubstituted or substituted by 1 or 2 F,    -   R₂₅(R₂₄)N—, wherein R₂₄ is H or (C₁-C₄)alkyl unsubstituted or        substituted by 1, 2 or 3 halo, R₂₅ is H or (C₁-C₄)alkyl        unsubstituted or substituted by 1, 2 or 3 halo,    -   OH

wherein n is 0, 1 or 2.

in particular R₁ is:

cycloalkenyl, wherein said cycloalkenyl is a partially unsaturatedmonocyclic ring containing 5 or 6 ring carbon atoms, and saidcycloalkenyl is unsubstituted or substituted by 1 or 2 R₃₃, wherein R₃₃is halo, preferably F, and wherein said cycloalkenyl or halo-substitutedcycloalkenyl is substituted by 0 or 1 R₁₅ substituents, preferably 1substituent, wherein R₁₅ is selected from:

-   -   h) (C₁-C₂)alkyl-O— unsubstituted or substituted by 1, 2 or 3        halo;    -   i) (C₁-C₂)alkyl unsubstituted or substituted by 1, 2 or 3 halo,    -   j) HOC(O)—(CH₂)_(n)—,    -   k) H₃C—C(O)(CH₂)_(n)—,    -   l) H₃C—O—C(O)(CH₂)_(n),    -   m) ═O, and    -   n) R₂₅(R₂₄)N—, H, wherein R₂₄ is H or (C₁-C₂)alkyl unsubstituted        or substituted by 1, 2 or 3 halo, R₂₅ is H or (C₁-C₂)alkyl        unsubstituted or substituted by 1, 2 or 3 halo,

n is 0 or 1,

wherein

-   -   the R₁₅ substituent a) to g) of said cycloalkenyl or        halo-substituted cycloalkenyl is not present on the ring atoms        adjacent to the ring atom to which the cycloalkenyl or        halo-substituted cycloalkenyl is joined to the remainder of the        molecule, and preferably, said cycloalkenyl or halo-substituted        cycloalkenyl is a 6 membered ring, with 1 R₁₅ substituent in the        ring para position relative to the remainder of the molecule;        and    -   said cycloalkenyl or halo-substituted cycloalkenyl is linked to        the remainder of the compound via a R₁ ring carbon atom which is        double bonded to an adjacent R₁ ring carbon atom;

or R₁ is heterocyclyl, wherein said heterocyclyl is a 5 or 6 memberedfully saturated or partially unsaturated group comprising ring carbonatoms and 1 or 2 ring heteroatoms independently selected from N, NH, Oand S, and wherein said heterocyclyl is unbridged or bridged, and saidbridge is 1 or 2 carbon atoms, wherein said heterocyclyl isunsubstituted or substituted by 1 or 2 R₃₃, wherein R₃₃ halo, ispreferably F, and wherein said heterocyclyl or halo-substitutedheterocyclyl is substituted by 0 or 1 substituents independentlyselected from R₁₅, R₁₆, R₁₇, R₁₈, R₁₉, R₂₀, R₂₂ and R₂₃, wherein saidR₁₅, R₁₆, R₁₇, R₁₈, R₁₉, R₂₀, R₂₂ and R₂₃ are independently selectedfrom:

-   -   i) (C₁-C₄)alkyl-O— unsubstituted or substituted by 1, 2 or 3        halo;    -   j) (C₁-C₄)alkyl unsubstituted or substituted by OH,        —O—(C₁-C₂)alkyl or 1, 2 or 3 halo,    -   k) HOC(O)—(CH₂)_(n)—,    -   l) H₃C—C(O)(CH₂)_(n)—,    -   m) H₃C—O—C(O)(CH₂)_(n),    -   n) ═O    -   o) R₂₅(R₂₄)N—, wherein R₂₄ is H, (C₁-C₂)alkyl unsubstituted or        substituted by 1, 2 or 3 halo, R₂₅ is H, (C₁-C₂)alkyl        unsubstituted or substituted by 1, 2 or 3 halo,    -   p) OH

wherein n is 0 or 1,

and wherein:

-   -   substituent a) to h) of said heterocyclyl or halo-substituted        heterocyclyl is not present on the ring atoms adjacent to the        ring atom to which the heterocyclyl or halo-substituted        heterocyclyl is joined to the remainder of the molecule, and        preferably, when said heterocyclyl or halo-substituted        heterocyclyl is a 6 membered ring, it has 0 or 1 substituent        selected from a) to h) in the meta or para position, preferably        para, relative to the remainder of the molecule; and    -   said heterocyclyl is linked to the remainder of the compound via        a R₁ ring nitrogen atom, or a R₁ ring carbon atom which is        double bonded to an adjacent ring atom;

or R₁ is heteroaryl, wherein said heteroaryl is a 5 or 6 membered fullyunsaturated monocyclic group comprising ring carbon atoms and 1 or 2ring heteroatoms independently selected from N, O and S, preferably N,wherein the total number of ring S atoms does not exceed 1, and thetotal number of ring O atoms does not exceed 1, wherein said heteroarylis unsubstituted or substituted by 1 or 2 substituents independentlyselected from R₂₁ and R₃₀, wherein R₂₁ and R₃₀ are independentlyselected from (C₁-C₂)alkyl, and said (C₁-C₂)alkyl is unsubstituted orsubstituted by 1, 2 or 3 halo, and wherein preferably, said alkyl orhalo-alkyl substituent is not present on the R₁ ring atoms adjacent tothe R₁ ring atom to which the heteroaryl is joined to the remainder ofthe molecule, and more preferably, when heteroaryl is a 6-membered ring,said alkyl or halo-alkyl substituent is in the ring para positionrelative to the rest of the molecule.

More particularly, R₁ is selected from:

R₃₃ is F;

R₁₅ is halo, azetidinyl or pyrrolidinyl, wherein said azetidinyl andpyrrolidinyl are linked to the rest of the molecule via the N atom, andare unsubstituted or substituted by 1 or 2 F;

R₁₆ is R₂₅(R₂₄)N—, wherein R₂₄ is H or (C₁-C₂)alkyl, R₂₅ is H or(C₁-C₂)alkyl unsubstituted or substituted by 1, 2 or 3 halo, inparticular F;

R₁₇ is halo

R₁₈ is halo;

R₁₉ is halo;

R₂₀ is halo;

R₂₁ is (C₁-C₂)alkyl;

R₂₂ and R₂₃ are each independently selected from:

-   -   (C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo,    -   HOC(O)—(CH₂)_(n)—,    -   H₃C—C(O)(CH₂)_(n)—,    -   (H₃C)₃C—O—C(O)(CH₂)_(n)—;    -   wherein n is 0, 1 or 2;

and

R₃₀ is CH₃.

Even more particularly, R₁ is selected from:

R₁₅ is F;

R₁₆ is R₂₅(R₂₄)N—;

R₁₇ is F;

R₁₈ is F;

R₁₉ is F;

R₂₀ is F;

R₂₁ is CH₃;

R₂₂ is CF₃, CHF₂CH₂, HOC(O)—CH₂—, H₃C—C(O)—, (H₃C)₃C—O—C(O)—;

R₂₃ is CF₃, CHF₂CH₂—, (H₃C)₃C—O—C(O)—;

R₂₄ is CH₃; and

R₂₅ is CHF₂CH₂—.

Preferably, R₁ is selected from:

In another embodiment, R₂ is the moiety:

R₆ is selected from:

-   -   H,    -   halo,    -   (C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo,    -   (C₃-C₅)cycloalkyl unsubstituted or substituted by 1, 2 or 3        halo,    -   —O—(C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo,    -   OH, and    -   CN;

R₆ is selected from H, halo, and (C₁-C₄)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo,

R₉ is selected from H, O—CH₃, OH, CN, CH₃ and halo;

R₂₈ is selected from:

-   -   SF₅,    -   H,    -   —C(O)H,    -   halo,    -   (C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo,    -   (C₁-C₄)alkynyl,    -   (C₁-C₄)alkenyl,    -   (C₃-C₅)cycloalkyl unsubstituted or substituted by 1, 2 or 3        halo, and    -   OCF₃;

and X is selected from C—R₇ and N, wherein R₇ is H or halo.

Particularly, R₂ is the moiety:

wherein

R₆ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₈ is selected from H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo;

R₉ is selected from H, O—CH₃, OH, CN, CH₃ and halo;

R₂₈ is selected from SF₅, halo, (C₁-C₄)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo and —C(O)H;

X is selected from C—R₇ and N; and

R₇ is selected from H and halo.

More particularly, R₂ is the moiety:

R₆ is selected from H, Cl, CH₃, F and Br;

R₈ is selected from H, Cl, F and CF₃;

R₉ is selected from H, CH₃ and Cl;

R₂₈ is selected from CF₃, CF₂H, —CH₂CH₃, Cl, SF₅, Br and —C(O)H;

X is selected from C—R₇ and N; and

R₇ is selected from H and F.

Even more particularly, R₂ is the moiety selected from:

preferably

In another embodiment, R₂₆ is H and R₂₇ is H.

In another embodiment, R₃ is (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 substituents independently selected from halo and OH, inparticular (C₁-C₂)alkyl unsubstituted or substituted by 1, 2 or 3substituents independently selected from halo and OH, preferably —CH₂CH₃or CH₃, more preferably —CH₂CH₃.

In another embodiment, Y is N and Y

is Y linked by a single bond.

In another embodiment, K

is K linked by a single bond, and K is selected from —CH₂—, —CH₂CH₂—,—NH— and a bond (to form a 5-membered ring

J is N, and A is a linker selected from —C(O)—, —S(O)—, —S(O)₂—, and

In particular,

K

is K linked by a single bond, K is —CH₂—, J is N, and A is a linkerselected from —C(O)—, —S(O)—, —S(O)₂—, and

In another embodiment, A is a linker selected from —C(O)— and —S(O)₂—,preferably —C(O)—.

In another embodiment, R₅ is independently selected from:

-   -   —(C₁-C₄)alkyl, preferably methyl,    -   and wherein two R₅ substituents on the same ring carbon atom may        join, together with the carbon atom to which they are attached,        to form a (C₃-C₄)cycloalkyl spiro ring or a 3 or 4-membered        heterocyclyl spiro ring, wherein said heterocyclyl spiro ring        contains ring carbon ring atoms and one ring heteroatom selected        from O, N and S,    -   when K        J is a carbon-nitrogen single bond, a R₅ substituent on K and on        the adjacent carbon atom may join to form ring C:

-   -   wherein ring C is a fused (C₃-C₆)cycloalkyl ring, in particular        a fused cyclobutyl ring, a fused (C₃-C₆)heterocyclyl ring or a        fused phenyl ring, wherein said fused (C₃-C₆)heterocyclyl ring        contains ring carbon atoms and one ring heteroatom selected from        O, N and S,    -   and wherein when K is —CH₂— and J is N, two R₅ substituents may        join to form a (C₁-C₃)alkylene bridge or a heteroalkylene        bridge, wherein said heteroalkylene bridge is one heteroatom        selected from N and O, or is —CH₂—O—CH₂—.

In particular, R₅ is independently selected from:

-   -   —(C₁-C₄)alkyl, preferably methyl,    -   when K        J is a carbon-nitrogen single bond, a R₅ substituent on K and on        the adjacent carbon atom may join to form ring C:

-   -   wherein ring C is a fused (C₃-C₆)cycloalkyl ring, in particular        a fused cyclobutyl ring, or a fused (C₃-C₆)heterocyclyl ring,        wherein said fused (C₃-C₆)heterocyclyl ring contains ring carbon        atoms and one ring heteroatom selected from O, N and S,    -   and wherein when K is —CH₂— and J is N, two R₅ substituents may        join to form a (C₁-C₃)alkylene bridge or a heteroalkylene        bridge, wherein said heteroalkylene bridge is one heteroatom        selected from N and O, or is —CH₂—O—CH₂—.

More particularly, R₅ is independently selected from:

-   -   —(C₁-C₂)alkyl, preferably methyl, and    -   when K        J is a carbon-nitrogen single bond, a R₅ substituent on K and on        the adjacent carbon atom may join to form ring C:

-   -   wherein ring C is a fused (C₃-C₄)cycloalkyl ring, in particular        a fused cyclobutyl ring.

Even more particularly, R₅ is independently selected from:

-   -   CH₃, and y is 1 or 2, and    -   when K        J is a carbon-nitrogen single bond, a R₅ substituent on K and on        the adjacent carbon atom may join to form ring C:

-   -   wherein ring C is a fused cyclobutyl ring.

In another embodiment, y is 0, 1, 2 or 3, preferably 0, 1, or 2.

In another embodiment, R₄ is selected from:

wherein

R₁₀ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents, —O—(C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents;

R₁₁ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents;

R₁₂ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents;

R₁₃ is selected from H, —S—CH₃, halo, (C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents; and

R₁₄ is selected from H, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents, O—(C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents, and cyclopropyl.

In particular, R₄ is selected from:

wherein

R₁₀ is selected from H, F, Cl, CH₃ and OCF₃;

R₁₁ is selected from H, Cl, F, and CH₃;

R₁₂ is selected from H, Cl and CH₃;

R₁₃ is selected from H, —S—CH₃ and CH₃; and

R₁₄ is selected from H, CH₃, —CH₂CH₃, cyclopropyl, —OCHF₂, OCF₃ and Cl.

More particularly, R₄ is selected from:

Even more particularly, R₄ is selected from:

preferably:

In another embodiment, formula (I) is formula 1a:

In another embodiment, the moiety:

is selected from

in particular:

Syntheses

There is also provided a process to manufacture a compound of formula(I) as described herein, in particular 1a, 1b, 1c, 1d, 1e, 1f, 1h or 1g,preferably 1a, more preferably 1g or 1 h, or a pharmaceuticallyacceptable salt thereof, as described herein.

There is further provided a intermediate compound, used in chemicalsynthesis of a compound of formula (I) as described herein, for exampleformula 1a, preferably 1g or 1h, or a pharmaceutically acceptable saltthereof, as described herein.

In another aspect, there is provided an intermediate compound, or aprocess comprising an intermediate compound, as described below.

In particular, there is provided a compound, or formula, as follows:

(sodium salt of the compound of Example 42)

A compound, or salt thereof, of formula A:

wherein R₁, R₂, R₃, R₂₆, R₂₇, R₅, y and Y are as defined herein.

A compound:

Intermediate AK

A compound, or salt thereof, of Formula B:

wherein R₁, R₂, R₃, R₂₆, R₂₇, R₅, y and Y are as defined herein, and PG₁is a protecting group. Suitable protecting groups are well-known to theskilled person, and include BOC.

A compound, or salt thereof, which is:

Intermediate AK step 1 product.

A compound, or salt thereof, of Formula C:

wherein R₁, R₃, R₅, y and Y are as defined herein, and PG₂ is aprotecting group. Suitable protecting groups are well-known to theskilled person. Such protecting groups PG₂ include BOC.

A compound, or salt thereof, which is:

Intermediate AF

A compound, or salt thereof, or a tautomer thereof, of Formula D:

wherein R₃, R₅, y and Y are as defined herein, and PG₃ is a protectinggroup. Suitable protecting groups are well-known to the skilled person.

A compound, or salt thereof, which is:

intermediate AF step 1 product

A compound, or salt thereof, which is:

Intermediate EH

There is also provided a process to make the compound:

-   -   comprising reacting the compound of formula:

Intermediate AK

-   -   with a compound of formula:

Intermediate DB

-   -   using coupling reagents and conditions known to the skilled        person, in particular HOAt, EDCI and DIPEA. Such reagents have        the advantages of being suitable for increased scale,        considering their availability and low cost, whilst enabling a        robust process with good yields. Protected or unprotected forms        can be used as known to the skilled person.

There is also provided a process to make the compound:

via the intermediates of formula:

intermediate AF, and/or

intermediate AF step 1 product, and/or

Intermediate EH.

Formulations

In another aspect, the present invention provides a pharmaceuticalcomposition comprising a compound of the present invention, or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier. In a further embodiment, the composition comprisesat least two pharmaceutically acceptable carriers, such as thosedescribed herein. The pharmaceutical composition can be formulated forparticular routes of administration such as oral administration,parenteral administration (e.g. by injection, infusion, transdermal ortopical administration), and rectal administration, in particular oraladministration. Topical administration may also pertain to inhalation orintranasal application. The pharmaceutical compositions of the presentinvention can be made up in a solid form (including, without limitation,capsules, tablets, pills, granules, powders or suppositories), or in aliquid form (including, without limitation, solutions, suspensions oremulsions). Tablets may be either film coated or enteric coatedaccording to methods known in the art. Typically, the pharmaceuticalcompositions are tablets or gelatin capsules comprising the activeingredient together with one or more of:

a) diluents, e.g., lactose, dextrose, sucrose, mannitol, sorbitol,cellulose and/or glycine;

b) lubricants, e.g., silica, talcum, stearic acid, its magnesium orcalcium salt and/or polyethyleneglycol; for tablets also

c) binders, e.g., magnesium aluminum silicate, starch paste, gelatin,tragacanth, methylcellulose, sodium carboxymethylcellulose and/orpolyvinylpyrrolidone; if desired

d) disintegrants, e.g., starches, agar, alginic acid or its sodium salt,or effervescent mixtures; and

e) absorbents, colorants, flavors and sweeteners.

Compounds intended for parenteral or oral administration can besolubilized using various methods including nano-suspensions, soliddispersions and liposomes (van Hoogevest P., Xiangli L., and Alfred F.“Drug delivery strategies for poorly water-soluble drugs: the industrialperspective” Expert Opinion on Drug Delivery 2011, 8(11), 1481-1500).

Solid dispersion technologies have been used to improve the dissolutioncharacteristics and bioavailability of orally administered drugs(Dhirendra K et al: ‘Solid dispersions: A review”, Pakistan Journal ofPharmaceutical Sciences, Faculty of Pharmacy, University of Karachi,Pakistan, vol. 22, no. 2. 30 April 200, pages 234-246).

Typical approaches to solubilize compounds for parenteral administrationare the optimization of the pH or the use of co-solvents (e.g. PEG300,PEG400, propylene glycol, or ethanol). If these approaches are, for anyreason, not feasible, the use of surfactants may be considered (e.g.Tween® 80 or Cremophor EL®). Cyclodextrins are established as safesolubilizing agents. Compounds with a high solubility in natural oils(e.g. propofol) may be solubilized in parenteral fat emulsions.

There is also provided a pharmaceutical composition comprising acompound of formula (I) as described herein, for example formula 1a,preferably 1g or 1h, or a pharmaceutically acceptable salt thereof, andone or more pharmaceutically acceptable carriers.

In another embodiment, there is provided a pharmaceutical compositionwhich is an amorphous solid dispersion comprising the compoundN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide,or a pharmaceutically acceptable salt thereof.

In a further embodiment, said pharmaceutical composition comprises thecompoundN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide,or a pharmaceutically acceptable salt thereof, and an amino methacrylatecopolymer, in particular Eudragit® E PO.

Uses

The compounds of formula (I) of the present invention in free form or inpharmaceutically acceptable salt form, exhibit valuable pharmacologicalproperties, e.g. WRN inhibiting properties, e.g. as indicated in vitroand in vivo tests as provided in the next sections, and are thereforeindicated for therapy, or for use as research chemicals, e.g. as achemical probe, and as tool compounds.

Also provided is a compound of formula (I), in particular 1a, 1b, 1c,1d, 1e, 1f, 1h or 1g, as described herein. Said compound can be used asa research chemical, for example a tool compound or chemical probe, inparticular for research on WRN. In another embodiment there is providedthe use of a compound of formula (I), in particular 1a, 1b, 1c, 1d, 1e,1f, 1h or 1g, as described herein, as a research chemical, for exampletool compound or chemical probe, in particular for research on WRN.

There is also provided a compound of formula (I) as described herein, inparticular 1a, 1b, 1c, 1d, 1e, 1f, 1h or 1g, preferably 1a, morepreferably 1g or 1h, or a pharmaceutically acceptable salt thereof, foruse in the treatment of cancer. Cancers that may be treated by WRNinhibition include cancers that are characterized as microsatelliteinstability-high (MSI-H) or mismatch repair deficient (dMMR). Inparticular, a compound of formula (I) as described herein, for exampleformula 1a, preferably 1g or 1h, or a pharmaceutically acceptable saltthereof, may be useful in the treatment of a cancer that ischaracterized as microsatellite instability-high (MSI-H) or mismatchrepair deficient (dMMR).

There is also provided a compound of formula (I) as described herein, inparticular 1a, 1b, 1c, 1d, 1e, 1f, 1h or 1g, preferably 1a, morepreferably 1g or 1h, or a pharmaceutically acceptable salt thereof, foruse as a medicament. In particular, said use is:

-   -   for the treatment of a disease that is treated by WRN        inhibition,    -   for the treatment of cancer,    -   for the treatment of cancer that is characterized as        microsatellite instability-high (MSI-H) or mismatch repair        deficient (dMMR),    -   for the treatment of cancer that is characterized as        microsatellite instability-high (MSI-H) or mismatch repair        deficient (dMMR), such as colorectal, gastric, prostate,        endometrial, adrenocortical, uterine, cervical, esophageal,        breast, kidney and ovarian cancer,    -   for the treatment of cancer that is characterized as        microsatellite instability-high (MSI-H) or mismatch repair        deficient (dMMR) is selected from colorectal, gastric, prostate        and endometrial cancer, or    -   for the treatment of cancer wherein the cancer characterized as        microsatellite instability-high (MSI-H) or mismatch repair        deficient (dMMR) is selected from uterine corpus endometrial        carcinoma, colon adenocarcinoma, stomach adenocarcinoma, rectal        adenocarcinoma, adrenocortical carcinoma, uterine        carcinosarcoma, cervical squamous cell carcinoma, endocervical        adenocarcinoma, esophageal carcinoma, breast carcinoma, kidney        renal clear cell carcinoma, prostate cancer and ovarian serous        cystadenocarcinoma.

There is also provided a method of:

-   -   modulating WRN activity in a subject, wherein the method        comprises administering to the subject a therapeutically        effective amount of the compound of formula (I) as described        herein, in particular 1a, 1b, 1c, 1d, 1e, 1f, 1h or 1g,        preferably 1a, more preferably 1g or 1h, or a pharmaceutically        acceptable salt thereof,    -   inhibiting WRN in a subject, wherein the method comprises        administering to the subject a therapeutically effective amount        of the compound of formula (I) as described herein, in        particular 1a, 1b, 1c, 1d, 1e, 1f, 1h or 1g, preferably 1a, more        preferably 1g or 1h, or a pharmaceutically acceptable salt        thereof,    -   treating a disorder or disease which can be treated by WRN        inhibition in a subject, comprising administering to the subject        a therapeutically effective amount of the compound of        formula (I) as described herein, in particular 1a, 1b, 1c, 1d,        1e, 1f, 1h or 1g, preferably 1a, more preferably 1g or 1h, or a        pharmaceutically acceptable salt thereof,    -   treating cancer in a subject, comprising administering to the        subject a therapeutically effective amount of the compound of        formula (I) as described herein, in particular 1a, 1b, 1c, 1d,        1e, 1f, 1h or 1g, preferably 1a, more preferably 1g or 1h, or a        pharmaceutically acceptable salt thereof,    -   treating cancer in a subject, comprising administering a        compound of formula (I) as described herein, in particular 1a,        1b, 1c, 1d, 1e, 1f, 1h or 1g, preferably 1a, more preferably 1g        or 1h, or a pharmaceutically acceptable salt thereof, wherein        the cancer is characterized as microsatellite instability-high        (MSI-H) or mismatch repair deficient (dMMR). In particular, the        cancer characterized as microsatellite instability-high (MSI-H)        or mismatch repair deficient (dMMR) is selected from colorectal,        gastric, prostate, endometrial, adrenocortical, uterine,        cervical, esophageal, breast, kidney and ovarian cancer. More        particularly, the cancer characterized as microsatellite        instability-high (MSI-H) or mismatch repair deficient (dMMR) is        selected from colorectal, gastric, prostate and endometrial        cancer. Examples include uterine corpus endometrial carcinoma,        colon adenocarcinoma, stomach adenocarcinoma, rectal        adenocarcinoma, adrenocortical carcinoma, uterine        carcinosarcoma, cervical squamous cell carcinoma, endocervical        adenocarcinoma, esophageal carcinoma, breast carcinoma, kidney        renal clear cell carcinoma, prostate cancer and ovarian serous        cystadenocarcinoma.

There is also provided the use of a compound of formula (I) as describedherein, in particular 1a, 1b, 1c, 1d, 1e, 1f, 1h or 1g, preferably 1a,more preferably 1g or 1h, or a pharmaceutically acceptable salt thereof:

-   -   in therapy,    -   in the manufacture of a medicament,    -   in the manufacture of a medicament for the treatment of cancer.        In particular, said cancer is characterized as microsatellite        instability-high (MSI-H) or mismatch repair deficient (dMMR),    -   in the manufacture of a medicament for treatment of a disease        which may be treated by WRN inhibition,

wherein in particular, the cancer is characterized by microsatelliteinstability-high (MSI-H) or mismatch repair deficient (dMMR), forexample colorectal, gastric, prostate, endometrial, adrenocortical,uterine, cervical, esophageal, breast, kidney and ovarian cancer, inparticular, colorectal, gastric, prostate or endometrial cancer, oruterine corpus endometrial carcinoma, colon adenocarcinoma, stomachadenocarcinoma, rectal adenocarcinoma, adrenocortical carcinoma, uterinecarcinosarcoma, cervical squamous cell carcinoma, endocervicaladenocarcinoma, esophageal carcinoma, breast carcinoma, kidney renalclear cell carcinoma and ovarian serous cystadenocarcinoma.

In some embodiments, the subject has or is identified as having amicrosatellite instable (MSI-H) cancer, e.g., in reference to a control,e.g., a normal, subject. In one embodiment, the subject has MSI-Hadvanced solid tumors, a colorectal cancer (CRC), endometrial, uterine,stomach or other MSI-H cancer. In some embodiments, the subject has acolorectal (CRC), endometrial or stomach cancer, which cancer has or isidentified as having a microsatellite instability (MSI-H), e.g., inreference to a control, e.g., a normal, subject. Such identificationtechniques are known in the art.

Forms

Depending on the choice of the starting materials and procedures, thecompounds can be present in the form of one of the possiblestereoisomers or as mixtures thereof, for example as pure opticalisomers, or as stereoisomer mixtures, such as racemates anddiastereoisomer mixtures, depending on the number of asymmetric carbonatoms. The present invention is meant to include all such possiblestereoisomers, including racemic mixtures, diasteriomeric mixtures andoptically pure forms. Optically active (R)- and (S)-stereoisomers may beprepared using chiral synthons or chiral reagents, or resolved usingconventional techniques. If the compound contains a double bond, thesubstituent may be E or Z configuration. If the compound contains adisubstituted cycloalkyl, the cycloalkyl substituent may have a cis- ortrans-configuration. All tautomeric forms are also intended to beincluded.

As used herein, the terms “salt” or “salts” refers to an acid additionor base addition salt of a compound of the present invention. “Salts”include in particular “pharmaceutical acceptable salts”. The term“pharmaceutically acceptable salts” refers to salts that retain thebiological effectiveness and properties of the compounds of thisinvention and, which typically are not biologically or otherwiseundesirable. In many cases, the compounds of the present invention arecapable of forming acid and/or base salts by virtue of the presence ofamino and/or carboxyl groups or groups similar thereto.

Pharmaceutically acceptable acid addition salts can be formed withinorganic acids and organic acids.

Inorganic acids from which salts can be derived include, for example,hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like.

Organic acids from which salts can be derived include, for example,acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid,malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid,benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid,toluenesulfonic acid, sulfosalicylic acid, and the like.

Pharmaceutically acceptable base addition salts can be formed withinorganic and organic bases.

Inorganic bases from which salts can be derived include, for example,ammonium salts and metals from columns I to XII of the periodic table.In certain embodiments, the salts are derived from sodium, potassium,ammonium, calcium, magnesium, iron, silver, zinc, and copper;particularly suitable salts include ammonium, potassium, sodium, calciumand magnesium salts.

Organic bases from which salts can be derived include, for example,primary, secondary, and tertiary amines, substituted amines includingnaturally occurring substituted amines, cyclic amines, basic ionexchange resins, and the like. Certain organic amines includeisopropylamine, benzathine, cholinate, diethanolamine, diethylamine,lysine, meglumine, piperazine and tromethamine.

In another aspect, the present invention provides compounds of thepresent invention in acetate, ascorbate, adipate, aspartate, benzoate,besylate, bromide/hydrobromide, bicarbonate/carbonate,bisulfate/sulfate, camphorsulfonate, caprate, chloride/hydrochloride,chlortheophyllonate, citrate, ethandisulfonate, fumarate, gluceptate,gluconate, glucuronate, glutamate, glutarate, glycolate, hippurate,hydroiodide/iodide, isethionate, lactate, lactobionate, laurylsulfate,malate, maleate, malonate, mandelate, mesylate, methylsulphate, mucate,naphthoate, napsylate, nicotinate, nitrate, octadecanoate, oleate,oxalate, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogenphosphate, polygalacturonate, propionate, sebacate, stearate, succinate,sulfosalicylate, sulfate, tartrate, tosylate trifenatate,trifluoroacetate or xinafoate salt form.

Any formula given herein is intended to represent unlabeled forms aswell as isotopically labeled forms of the compounds, in addition to thedeuteration specifically claimed in formula (I). Isotopically labeledcompounds have structures depicted by the formulae given herein exceptthat one or more atoms are replaced by an atom having a selected atomicmass or mass number. Isotopes that can be incorporated into compounds ofthe invention include, for example, isotopes of hydrogen.

Further, incorporation of certain isotopes, particularly deuterium(i.e., ²H or D) may afford certain therapeutic advantages resulting fromgreater metabolic stability, for example increased in vivo half-life orreduced dosage requirements or an improvement in therapeutic index ortolerability. It is understood that deuterium in this context isregarded as a substituent of a compound of the present invention. Theconcentration of deuterium, may be defined by the isotopic enrichmentfactor. The term “isotopic enrichment factor” as used herein means theratio between the isotopic abundance and the natural abundance of aspecified isotope. If a substituent in a compound of this invention isdenoted as being deuterium, such compound has an isotopic enrichmentfactor for each designated deuterium atom of at least 3500 (52.5%deuterium incorporation at each designated deuterium atom), at least4000 (60% deuterium incorporation), at least 4500 (67.5% deuteriumincorporation), at least 5000 (75% deuterium incorporation), at least5500 (82.5% deuterium incorporation), at least 6000 (90% deuteriumincorporation), at least 6333.3 (95% deuterium incorporation), at least6466.7 (97% deuterium incorporation), at least 6600 (99% deuteriumincorporation), or at least 6633.3 (99.5% deuterium incorporation). Itshould be understood that the term “isotopic enrichment factor” can beapplied to any isotope in the same manner as described for deuterium.

Other examples of isotopes that can be incorporated into compounds ofthe invention include isotopes of hydrogen, carbon, nitrogen, oxygen,phosphorous, fluorine, and chlorine, such as ³H, ¹¹C, ¹³C, ¹⁴C, ¹⁵N, ¹⁸F³¹P, ³²P, ³⁵S, ³⁶Cl, ¹²³I, ¹²⁴I, ¹²⁵I respectively. Accordingly itshould be understood that the invention includes compounds thatincorporate one or more of any of the aforementioned isotopes, includingfor example, radioactive isotopes, such as ³H and ¹⁴C, or those intowhich non-radioactive isotopes, such as ²H and ¹³C are present. Suchisotopically labelled compounds are useful in metabolic studies (with¹⁴C), reaction kinetic studies (with, for example ²H or ³H), detectionor imaging techniques, such as positron emission tomography (PET) orsingle-photon emission computed tomography (SPECT) including drug orsubstrate tissue distribution assays, or in radioactive treatment ofpatients. In particular, an ¹⁸F or labeled compound may be particularlydesirable for PET or SPECT studies. Isotopically-labeled compounds ofthe present invention can generally be prepared by conventionaltechniques known to those skilled in the art or by processes analogousto those described in the accompanying Examples and Preparations usingan appropriate isotopically-labeled reagents in place of the non-labeledreagent previously employed.

Deuterated compounds of formula (I) include example 43, a deuteratedform of example 42:

and deuterated forms of the compounds of:

Example 86

Example 50

Example 57

and example 47:

Definitions

A ‘compound of the present invention’ or a ‘compound of formula (I)’ ora ‘compound of formula 1a’, or 1g or h etc, includes a zwitterionthereof, a non-zwitterion thereof (non-charged form), or apharmaceutically acceptable salt of said zwitterionic ornon-zwitterionic form thereof.

‘zwitterion’ or ‘zwitterionic form’ means a compound containing bothpositive and negatively charged functional groups.

For example, the compound of formula (I) described herein can includethe following forms, wherein R₄ is the zwitterionic form (c) ornon-zwitterionic form (d),

or a mixture thereof.

The compound of formula (I) described herein can also include thefollowing forms, wherein R₄ is the zwitterionic form (a) or (b) or thenon-zwitterionic form (e),

or a mixture of two thereof, or a mixture of all three thereof.

halo means fluoro, chloro or bromo, particularly fluoro or chloro.

Alkyl, and alkoxy groups, containing the requisite number of carbonatoms, can be unbranched or branched. Examples of alkyl include, but arenot limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl,sec-butyl and t-butyl. Examples of alkoxy include methoxy, ethoxy,n-propoxy, i-propoxy, n-butoxy, i-butoxy, sec-butoxy and t-butoxy.

‘═O’ means an oxo substituent.

When R₁ is substituted or unsubstituted cycloalkenyl, said cycloalkenylincludes, but is not limited to, groups such as cyclohexenyl, inparticular cyclohex-1-en-1-yl.

When R₁ is substituted or unsubstituted heterocyclyl, said heterocyclylincludes, but is not limited to, groups such as morpholinyl,piperidinyl, pyrrolidinyl, 6-oxa-3-azabicyclo[3.1.1]heptan-3-yl,5,6-dihydro-1,4-dioxin-2-yl, dihydropyranyl, in particular3,4-dihydro-2H-pyran-6-yl, 5,6-dihydro-2H-pyran-3-yl and3,6-dihydro-2H-pyran-4-yl, piperazinyl, tetrahydropyridinyl, such as1,4,5,6-tetrahydropyridin-3-yl and 1,2,3,6-tetrahydropyridin-4-yl anddihydropyridinyl, such as 3,6-dihydropyridinyl.

When R₁ is substituted or unsubstituted heteroaryl, said heteroarylincludes, but is not limited to, groups such as pyridinyl, in particularpyridin-3-yl.

The term “cancer” refers to a disease characterized by the rapid anduncontrolled growth of aberrant cells. Cancer cells can spread locallyor through the bloodstream and lymphatic system to other parts of thebody. Examples of various cancers are described herein and include butare not limited to colorectal, gastric, endometrial, prostate,adrenocortical, uterine, cervical, esophageal, breast, kidney, ovariancancer and the like.

The terms “tumor” and “cancer” are used interchangeably herein, e.g.,both terms encompass solid and liquid, e.g., diffuse or circulating,tumors. As used herein, the term “cancer” or “tumor” includespremalignant, as well as malignant cancers and tumors.

‘WRN inhibitor’ or ‘WRN helicase inhibitor’ as used herein means acompound that inhibits Werner Syndrome RecQ DNA helicase (WRN). The term“WRN” as used herein refers to the protein of Werner Syndrome RecQ DNAhelicase. The term “WRN” includes mutants, fragments, variants,isoforms, and homologs of full-length wild-type WRN. In one embodiment,the protein is encoded by the WRN gene (Entrez gene ID 7486; Ensembl IDENSG00000165392). Exemplary WRN sequences are available at the Uniprotdatabase under accession number Q14191.

‘disease or condition mediated by WRN’ includes a disease or condition,such as cancer, which is treated by WRN inhibition. In particular thiscan include cancers characterized as microsatellite instability-high(MSI-H) or mismatch repair deficient (dMMR).

‘microsatellite unstable cancer’, microsatellite instability-highcancer’, ‘microsatellite high cancer’ and ‘MSI-high cancer’ ‘MSI^(hi)’and ‘MSI-H’ when used herein, are used interchangeably, and describecancers that have a high number of alterations in the length of simplerepetitive genomic sequences within microsatellites.

The determination of MSI-H or dMMR tumor status for patients can beperformed using, e.g., polymerase chain reaction (PCR) tests for MSI-Hstatus or immunohistochemistry (IHC) tests for dMMR. Methods foridentification of MSI-H or dMMR tumor status are described, e.g., inRyan et al. Crit Rev Oncol Hematol. 2017; 116:38-57; Dietmaier andHofstadter. Lab Invest 2001, 81:1453-1456; and Kawakami et al. CurrTreat Options Oncol. 2015; 16(7): 30).

Microsatellite instability can be found in colorectal cancer, gastriccancer and endometrial cancer in particular, but also in adrenocortical,uterine, cervical, esophageal, breast, kidney, prostate and ovariancancers. Examples of microsatellite high cancers include uterine corpusendometrial carcinoma, colon adenocarcinoma, stomach adenocarcinoma,rectal adenocarcinoma, adrenocortical carcinoma, uterine carcinosarcoma,cervical squamous cell carcinoma, endocervical adenocarcinoma,esophageal carcinoma, breast carcinoma, kidney renal clear cellcarcinoma and ovarian serous cystadenocarcinoma.

A cancer that has “defective mismatch repair” (dMMR) or “dMMR character”includes cancer types associated with documented MLH1, PMS2, MSH2, MSH3,MSH6, MLH3, and PMS1 mutations or epigenetic silencing, microsatellitefragile sites, or other gene inactivation mechanisms, including but notlimited to cancers of the lung, breast, kidney, large intestine, ovary,prostate, upper aerodigestive tract, stomach, endometrium, liver,pancreas, haematopoietic and lymphoid tissue, skin, thyroid, pleura,autonomic ganglia, central nervous system, soft tissue, pediatricrhabdoid sarcomas, melanomas and other cancers. A cell or cancer with“defective” mismatch repair has a significantly reduced (e.g., at leastabout 25%, 30%, 40%, 50%, 60%, 70%, 80% or 90% decrease) amount ofmismatch repair. In some cases, a cell or cancer which is defective inmismatch repair will perform no mismatch repair.

As used herein, the term “pharmaceutical composition” refers to acompound of the invention, or a pharmaceutically acceptable saltthereof, together with at least one pharmaceutically acceptable carrier,in a form suitable for oral or parenteral administration.

As used herein, the term “pharmaceutically acceptable carrier” refers toa substance useful in the preparation or use of a pharmaceuticalcomposition and includes, for example, suitable diluents, solvents,dispersion media, surfactants, antioxidants, preservatives, isotonicagents, buffering agents, emulsifiers, absorption delaying agents,salts, drug stabilizers, binders, excipients, disintegration agents,lubricants, wetting agents, sweetening agents, flavoring agents, dyes,and combinations thereof, as would be known to those skilled in the art(see, for example, Remington The Science and Practice of Pharmacy, 22ndEd. Pharmaceutical Press, 2013, pp. 1049-1070).

The terms “synthetic lethality,” and “synthetic lethal” are used torefer to reduced cell viability and/or a reduced rate of cellproliferation caused by a combination of mutations or approaches tocause loss of function (e.g., RNA interference or protein functioninhibition) in two or more genes but not by the loss of function of onlyone of these genes.

The term “a therapeutically effective amount” of a compound of thepresent invention refers to an amount of the compound of the presentinvention that will elicit the biological or medical response of asubject, for example, reduction or inhibition of an enzyme or a proteinactivity, or ameliorate symptoms, alleviate conditions, slow or delaydisease progression, or prevent a disease, etc.

In one embodiment, the term “a therapeutically effective amount” refersto the amount of the compound of the present invention that, whenadministered to a subject, is effective to (1) at least partiallyalleviate, prevent and/or ameliorate a condition, or a disorder or adisease (i) mediated by WRN, or (ii) associated with WRN activity, or(iii) characterized by activity (normal or abnormal) of WRN; or (2)reduce or inhibit the activity of WRN.

In another embodiment, the term “a therapeutically effective amount”refers to the amount of the compound of the present invention that, whenadministered to a cell, or a tissue, or a non-cellular biologicalmaterial, or a medium, is effective to at least partially reducing orinhibiting the activity of WRN, or reducing WRN protein levels.

As used herein, the term “subject” refers to primates (e.g., humans,male or female), dogs, rabbits, guinea pigs, pigs, rats and mice. Incertain embodiments, the subject is a primate, a rat or a mouse. In yetother embodiments, the subject is a human.

As used herein, the term “inhibit”, “inhibition” or “inhibiting” refersto the reduction or suppression of a given condition, symptom, ordisorder, or disease, or a significant decrease in the baseline activityof a biological activity or process.

As used herein, the term “treat”, “treating” or “treatment” of anydisease or disorder refers to alleviating or ameliorating the disease ordisorder (i.e., slowing or arresting the development of the disease orat least one of the clinical symptoms thereof); or alleviating orameliorating at least one physical parameter or biomarker associatedwith the disease or disorder, including those which may not bediscernible to the patient.

As used herein, the term “prevent”, “preventing” or “prevention” of anydisease or disorder refers to the prophylactic treatment of the diseaseor disorder; or delaying the onset or progression of the disease ordisorder.

As used herein, a subject is “in need of” a treatment if such subjectwould benefit biologically, medically or in quality of life from suchtreatment.

As used herein, the term “a,” “an,” “the” and similar terms used in thecontext of the present invention (especially in the context of theclaims) are to be construed to cover both the singular and plural unlessotherwise indicated herein or clearly contradicted by the context.

‘May join’ means joins or does not join.

‘May be replaced by deuterium’ means is replaced by deuterium, or is notreplaced by deuterium.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although methods and materialssimilar or equivalent to those described herein can be used in thepractice or testing of the present invention, suitable methods andmaterials are described below. All publications, patent applications,patents, and other references mentioned herein are incorporated byreference in their entirety. In addition, the materials, methods, andexamples are illustrative only and not intended to be limiting. Allmethods described herein can be performed in any suitable order unlessotherwise indicated herein or otherwise clearly contradicted by context.The use of any and all examples, or exemplary language (e.g. “such as”)provided herein is intended merely to better illuminate the inventionand does not pose a limitation on the scope of the invention otherwiseclaimed.

Isomeric Forms

Any asymmetric atom (e.g., carbon or the like) of the compound(s) of thepresent invention can be present in racemic or enantiomericallyenriched, for example the (R)-, (S)- or (R,S)-configuration. In certainembodiments, each asymmetric atom has at least 50% enantiomeric excess,at least 60% enantiomeric excess, at least 70% enantiomeric excess, atleast 80% enantiomeric excess, at least 90% enantiomeric excess, atleast 95% enantiomeric excess, or at least 99% enantiomeric excess inthe (R)- or (S)-configuration.

Substituents at atoms with unsaturated double bonds may, if possible, bepresent in cis-(Z)- or trans-(E)-form.

Accordingly, as used herein a compound of the present invention can bein the form of one of the possible stereoisomers, rotamers,atropisomers, tautomers or mixtures thereof, for example, assubstantially pure geometric (cis or trans) stereoisomers,diastereomers, optical isomers (antipodes), racemates or mixturesthereof.

Any resulting mixtures of stereoisomers can be separated on the basis ofthe physicochemical differences of the constituents, into the pure orsubstantially pure geometric or optical isomers, diastereomers,racemates, for example, by chromatography and/or fractionalcrystallization.

Any resulting racemates of compounds of the present invention or ofintermediates can be resolved into the optical antipodes by knownmethods, e.g., by separation of the diastereomeric salts thereof,obtained with an optically active acid or base, and liberating theoptically active acidic or basic compound. In particular, a basic moietymay thus be employed to resolve the compounds of the present inventioninto their optical antipodes, e.g., by fractional crystallization of asalt formed with an optically active acid, e.g., tartaric acid,dibenzoyl tartaric acid, diacetyl tartaric acid, di-O,O′-p-toluoyltartaric acid, mandelic acid, malic acid or camphor-10-sulfonic acid.Racemic compounds of the present invention or racemic intermediates canalso be resolved by chiral chromatography, e.g., high pressure liquidchromatography (HPLC) using a chiral adsorbent.

Compounds of the invention, i.e. compounds of formula (I) that containgroups capable of acting as donors and/or acceptors for hydrogen bondsmay be capable of forming co-crystals with suitable co-crystal formers.These co-crystals may be prepared from compounds of formula (I) by knownco-crystal forming procedures. Such procedures include grinding,heating, co-subliming, co-melting, or contacting in solution compoundsof formula (I) with the co-crystal former under crystallizationconditions and isolating co-crystals thereby formed. Suitable co-crystalformers include those described in WO 2004/078163. Hence the inventionfurther provides co-crystals comprising a compound of formula (I).

Furthermore, the compounds of the present invention, including theirsalts, can also be obtained in the form of their hydrates, or includeother solvents used for their crystallization.

The compounds of the present invention may inherently or by design formsolvates with pharmaceutically acceptable solvents (including water);therefore, it is intended that the invention embrace both solvated andunsolvated forms. The term “solvate” refers to a molecular complex of acompound of the present invention (including pharmaceutically acceptablesalts thereof) with one or more solvent molecules. Such solventmolecules are those commonly used in the pharmaceutical art, which areknown to be innocuous to the recipient, e.g., water, ethanol, and thelike. The term “hydrate” refers to the complex where the solventmolecule is water.

Dosage Forms

The pharmaceutical composition or combination of the present inventionmay, for example, be in unit dosage of about 1-1000 mg of activeingredient(s) for a subject of about 50-70 kg.

Combinations

“Combination” refers to either a fixed combination in one dosage unitform, or a combined administration where a compound of formula (I), or apharmaceutically acceptable salt thereof, and a combination partner(e.g. another drug as explained below, also referred to as “therapeuticagent” or “co-agent”) may be administered independently at the same timeor separately within time intervals, especially where these timeintervals allow that the combination partners show a cooperative, e.g.synergistic effect. The single components may be packaged in a kit orseparately. One or both of the components (e.g., powders or liquids) maybe reconstituted or diluted to a desired dose prior to administration.The terms “co-administration” or “combined administration” or the likeas utilized herein are meant to encompass administration of the selectedcombination partner to a single subject in need thereof (e.g. apatient), and are intended to include treatment regimens in which theagents are not necessarily administered by the same route ofadministration or at the same time. The term “pharmaceuticalcombination” as used herein means a product that results from the mixingor combining of more than one therapeutic agent and includes both fixedand non-fixed combinations of the therapeutic agents. The term “fixedcombination” means that the therapeutic agents, e.g. a compound of thepresent invention and a combination partner, are both administered to apatient simultaneously in the form of a single entity or dosage.

The term “non-fixed combination” means that the therapeutic agents, e.g.a compound of the present invention and a combination partner, are bothadministered to a patient as separate entities either simultaneously,concurrently or sequentially with no specific time limits, wherein suchadministration provides therapeutically effective levels of the twocompounds in the body of the patient. The latter also applies tococktail therapy, e.g. the administration of three or more therapeuticagents.

The combinations described herein can include a compound of formula (I)and one or more additional therapeutic agents, e.g., one or moreanti-cancer agents, cytotoxic or cytostatic agents, hormone treatment,vaccines, and/or other immunotherapies. In other embodiments, thecombination is further administered or used in combination with othertherapeutic treatment modalities, including surgery, radiation,cryosurgery, and/or thermotherapy. Such combination therapies mayadvantageously utilize lower dosages of the administered therapeuticagents, thus avoiding possible toxicities or complications associatedwith the treatment.

There is also provided a combination comprising a compound of formula(I) as described herein, in particular 1a, 1b, 1c, 1d, 1e, 1f, 1h or 1g,preferably 1a, more preferably 1g or 1 h, or a pharmaceuticallyacceptable salt thereof, as described herein, and one or more additionaltherapeutically active agents. The additional therapeutic agent is, forexample, a chemical compound, peptide, antibody, antibody fragment ornucleic acid, which is therapeutically active or enhances thetherapeutic activity when administered to a patient in combination witha compound of the present disclosure. In particular, an additionaltherapeutically active agent is:

-   -   an anti-cancer agent,    -   a chemotherapy,    -   a chemotherapy selected from anastrozole (Arimidex®),        bicalutamide (Casodex®), bleomycin sulfate (Blenoxane®),        busulfan (Myleran®), busulfan injection (Busulfex®),        capecitabine (Xeloda®),        N4-pentoxycarbonyl-5-deoxy-5-fluorocytidine, carboplatin        (Paraplatin®), carmustine (BiCNU®), chlorambucil (Leukeran®),        cisplatin (Platinol®), cladribine (Leustatin®), cyclophosphamide        (Cytoxan® or Neosar®), cytarabine, cytosine arabinoside        (Cytosar-U®), cytarabine liposome injection (DepoCyt®),        dacarbazine (DTIC-Dome®), dactinomycin (Actinomycin D,        Cosmegan), daunorubicin hydrochloride (Cerubidine®),        daunorubicin citrate liposome injection (DaunoXome®),        dexamethasone, docetaxel (Taxotere®), doxorubicin hydrochloride        (Adriamycin®, Rubex®), etoposide (Vepesid®), fludarabine        phosphate (Fludara®), 5-fluorouracil (Adrucil®, Efudex®),        flutamide (Eulexin®), tezacitibine, Gemcitabine        (difluorodeoxycitidine), hydroxyurea (Hydrea®), Idarubicin        (Idamycin®), ifosfamide (IFEX®), irinotecan (Camptosar®),        L-asparaginase (ELSPAR®), leucovorin calcium, melphalan        (Alkeran®), 6-mercaptopurine (Purinethol®), methotrexate        (Folex®), mitoxantrone (Novantrone®), mylotarg, paclitaxel        (Taxol®), phoenix (Yttrium90/MX-DTPA), pentostatin, polifeprosan        20 with carmustine implant (Gliadel®), tamoxifen citrate        (Nolvadex®), teniposide (Vumon®), 6-thioguanine, thiotepa,        tirapazamine (Tirazone®), topotecan hydrochloride for injection        (Hycamptin®), vinblastine (Velban®), vincristine (Oncovin®), and        vinorelbine (Navelbine®), in particular fluorouracil (5-FU) and        irinotecan (Camptosar®).    -   a PD-1 inhibitor,    -   an anti-PD-1 antibody molecule, or    -   a PD-1 inhibitor selected from PDR001 (Novartis), Nivolumab        (Bristol-Myers Squibb), Pembrolizumab (Merck & Co), Pidilizumab        (CureTech), MEDI0680 (Medimmune), Cemiplimab (REGN2810,        Regeneron), Dostarlimab (TSR-042, Tesaro), PF-06801591 (Pfizer),        Tislelizumab (BGB-A317, Beigene), BGB-108 (Beigene), INCSHR1210        (Incyte), Balstilimab (AGEN2035, Agenus), Sintilimab (InnoVent),        Toripalimab (Shanghai Junshi Bioscience), Camrelizumab (Jiangsu        Hengrui Medicine Co.), and AMP-224 (Amplimmune), Penpulimab        (Akeso Biopharma Inc), Zimberelimab (Arcus Biosciences Inc) and        Prolgolimab (Biocad Ltd), in particular PDR001, more        particularly Tislelizumab (BGB-A317, Beigene).

In a further embodiment, the additional therapeutically active agent isthe chemotherapy irinotecan (Camptosar®).

In another embodiment, the additional therapeutically active agent is aninhibitor of PD-1, e.g., human PD-1. In another embodiment, theimmunomodulator is an inhibitor of PD-L1, e.g., human PD-L1. In oneembodiment, the inhibitor of PD-1 or PD-L1 is an antibody molecule toPD-1 or PD-L1. In another embodiment, the additional therapeuticallyactive agent is an anti-PD-1 antibody molecule.

In a further embodiment, the PD-1 inhibitor is an anti-PD-1 antibodymolecule as described in US 2015/0210769, published on Jul. 30, 2015,entitled “Antibody Molecules to PD-1 and Uses Thereof,” incorporated byreference in its entirety.

In another embodiment, there is provided a combination of a compound offormula (I) or a pharmaceutically acceptable salt thereof, and achemotherapy, and a PD-1 inhibitor. In particular, the chemotherapy andPD-1 inhibitor are selected from those described above. Moreparticularly, the chemotherapy is irinotecan (Camptosar®) and the PD-1inhibitor is PDR01 or Tislelizumab. Tislelizumab can have a heavy chainof SEQ ID NO: 3 and a light chain of SEQ ID NO: 4. In some embodiments,the anti-PD-1 antibody is dosed at 100 mg per week. In some embodiments,tislelizumab and is dosed at 300 mg IV on day 1 of each 28 day cycle. Insome embodiments, tislelizumab can be dosed at 500 mg once every four(4) weeks.

In another embodiment, the anti-PD-1 antibody molecule, e.g.,tislelizumab, and comprises a heavy chain and/or light chain, VH, VL,HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of the following:

AMINO ACID SEQUENCE Heavy QVQLQESGPGLVKPSETLSL SEQ ID NO: 3 ChainTCTVSGFSLTSYGVHWIRQP PGKGLEWIGVIYADGSTNYN PSLKSRVTISKDTSKNQVSLKLSSVTAADTAVYYCARAYG NYWYIDVWGQGTTVTVSSAS TKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKY GPPCPPCPAPPVAGGPSVFL FPPKPKDTLMISRTPEVTCWVAVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVV SVLTVVHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQP REPQVYTLPPSQEEMTKNQV SLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS FFLYSKLTVDKSRWQEGNVF SCSVMHEALHNHYTQKSLSL SLGKLight DIVMTQSPDSLAVSLGERAT SEQ ID NO: 4 Chain INCKSSESVSNDVAWYQQKPGQPPKLLINYAFHRFTGVPD RFSGSGYGTDFTLTISSLQA EDVAVYYCHQAYSSPYTFGQGTKLEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFY PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQG LSSPVTKSFNRGEC VariableQVQLQESGPGLVKPSETLSL SEQ ID NO: 5 Heavy TCTVSGFSLTSYGVHWIRQP ChainPGKGLEWIGVIYADGSTNYN PSLKSRVTISKDTSKNQVSL KLSSVTAADTAVYYCARAYGNYWYIDVWGQGTTVTVSS  Variable DIVMTQSPDSLAVSLGERAT SEQ ID NO: 6 LightINCKSSESVSNDVAWYQQKP Chain GQPPKLLINYAFHRFTGVPD RFSGSGYGTDFTLTISSLQAEDVAVYYCHQAYSSPYTFGQ GTKLEIK  HCDR1 GFSLTSYGVH SEQ ID NO: 7 HCDR2VIYADGSTNYNPSLKS SEQ ID NO: 8 HCDR3 ARAYGNYWYIDV SEQ ID NO: 9 LCDR1KSSESVSNDVA SEQ ID NO: 10 LCDR2 YAFHRFT SEQ ID NO: 11 LCDR3 HQAYSSPYTSEQ ID NO: 12

In some embodiments, the PD-1 inhibitor comprises the HCDRs and LCDRs oftislelizumab as set forth in SEQ ID NOs: 7-12.

In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) isadministered at a flat dose of between about 100 mg to about 600 mg. Insome embodiments, the PD-1 inhibitor is administered at a dose ofbetween about 100 mg to about 500 mg. In some embodiments, the PD-1inhibitor is administered at a dose of between about 100 mg to about 400mg. In some embodiments, the PD-1 inhibitor is administered at a dose ofbetween about 100 mg to about 300 mg. In some embodiments, the PD-1inhibitor is administered at a dose of between about 100 mg to about 200mg. In some embodiments, the PD-1 inhibitor is administered at a dose ofbetween about 200 mg to about 600 mg. In some embodiments, the PD-1inhibitor is administered at a dose of between about 200 mg to about 500mg. In some embodiments, the PD-1 inhibitor is administered at a dose ofbetween about 200 mg to about 400 mg. In some embodiments, the PD-1inhibitor is administered at a dose of between about 200 mg to about 300mg. In some embodiments, the PD-1 inhibitor is administered at a dose ofbetween about 300 mg to about 600 mg. In some embodiments, the PD-1inhibitor is administered at a dose of between about 300 mg to about 500mg. In some embodiments, the PD-1 inhibitor is administered at a dose ofbetween about 300 mg to about 400 mg. In some embodiments, the PD-1inhibitor is administered at a dose of between about 400 mg to about 600mg. In some embodiments, the PD-1 inhibitor is administered at a dose ofbetween about 400 mg to about 500 mg. In some embodiments, the PD-1inhibitor is administered at a dose of between about 500 mg to about 600mg. In some embodiments, the PD-1 inhibitor is administered at a dose ofbetween about 600 mg to about 700 mg. In some embodiments, the PD-1inhibitor is administered at a dose of between about 700 mg to about 800mg. In some embodiments, the PD-1 inhibitor is administered at a dose ofbetween about 800 mg to about 900 mg. In some embodiments, the PD-1inhibitor is administered at a dose of between about 900 mg to about1000 mg.

In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) isadministered at a flat dose of about 100 mg. In some embodiments, thePD-1 inhibitor is administered at a dose of about 200 mg. In someembodiments, the PD-1 inhibitor is administered at a dose of about 300mg. In some embodiments, the PD-1 inhibitor is administered at a dose ofabout 400 mg. In some embodiments, the PD-1 inhibitor is administered ata dose of about 500 mg. In some embodiments, the PD-1 inhibitor isadministered at a dose of about 600 mg. In some embodiments, the PD-1inhibitor is administered at a dose of about 700 mg. In someembodiments, the PD-1 inhibitor is administered at a dose of about 800mg. In some embodiments, the PD-1 inhibitor is administered at a dose ofabout 900 mg. In some embodiments, the PD-1 inhibitor is administered ata dose of about 1000 mg.

In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) isadministered once every ten weeks. In some embodiments, the PD-1inhibitor is administered once every nine weeks. In some embodiments,the PD-1 inhibitor is administered once every eight weeks. In someembodiments, the PD-1 inhibitor is administered once every seven weeks.In some embodiments, the PD-1 inhibitor is administered once every sixweeks. In some embodiments, the PD-1 inhibitor is administered onceevery five weeks. In some embodiments, the PD-1 inhibitor isadministered once every four weeks. In some embodiments, the PD-1inhibitor is administered once every three weeks. In some embodiments,the PD-1 inhibitor is administered once every two weeks. In someembodiments, the PD-1 inhibitor is administered once every week.

In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) isadministered intravenously.

In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) isadministered over a period of about 20 minutes to 40 minutes (e.g.,about 30 minutes). In some embodiments, the PD-1 inhibitor isadministered over a period of about 30 minutes. In some embodiments, thePD-1 inhibitor is administered over a period of about an hour. In someembodiments, the PD-1 inhibitor is administered over a period of abouttwo hours. In some embodiments, the PD-1 inhibitor is administered overa period of about three hours. In some embodiments, the PD-1 inhibitoris administered over a period of about four hours. In some embodiments,the PD-1 inhibitor is administered over a period of about five hours. Insome embodiments, the PD-1 inhibitor is administered over a period ofabout six hours.

In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) isadministered at a dose between about 300 mg to about 500 mg (e.g., about400 mg), intravenously, once every four weeks. In some embodiments, thePD-1 inhibitor is administered at a dose between about 200 mg to about400 mg (e.g., about 300 mg), intravenously, once every three weeks. Insome embodiments, tislelizumab is administered at a dose of 400 mg, onceevery four weeks. In some embodiments, tislelizumab is administered at adose of 300 mg, once every three weeks.

In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) isadministered at a dose between about 300 mg to about 500 mg (e.g., about400 mg), intravenously, over a period of about 20 minutes to about 40minutes (e.g., about 30 minutes), once every two weeks. In someembodiments, the PD-1 inhibitor is administered at a dose between about200 mg to about 400 mg (e.g., about 300 mg), intravenously, over aperiod of about 20 minutes to about 40 minutes (e.g., about 30 minutes),once every three weeks.

In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) isadministered at a dose of about 100 mg per week. For example, if a10-week dose is given to a patient, then the PD-1 inhibitor (e.g.,tislelizumab) can be given at 1000 mg. If a 9-week dose is given, thenthe PD-1 inhibitor (e.g., tislelizumab) can be given at 900 mg. If an8-week dose is given, then the PD-1 inhibitor (e.g., tislelizumab) canbe given at 800 mg. If a 7-week dose is given, then the PD-1 inhibitor(e.g., tislelizumab) can be given at 700 mg. If a 6-week dose is given,then the PD-1 inhibitor (e.g., tislelizumab) can be given at 600 mg. Ifa 5-week dose is given, then the PD-1 inhibitor (e.g., tislelizumab) canbe given at 500 mg. If a 4-week dose is given, then the PD-1 inhibitor(e.g., tislelizumab) can be given at 400 mg. If a 3-week dose is given,then the PD-1 inhibitor (e.g., tislelizumab) can be given at 300 mg. Ifa 2-week dose is given, then the PD-1 inhibitor (e.g., tislelizumab) canbe given at 200 mg. If a 1-week dose is given, then the PD-1 inhibitor(e.g., tislelizumab) can be given at 100 mg.

For example, if an anti-PD-1 antibody, such as tislelizumab is used, itcan be administered at a dose of 200 mg as an intravenous infusion, onceevery three week. Alternatively, tislelizumab can be administered at adose of 300 mg as an intravenous infusion, once every four weeks. If ananti-PD-1 antibody, such as tislelizumab is used, it can be administeredat a dose of 300 mg as an intravenous infusion, once every three week.Alternatively, tislelizumab can be administered at a dose of 400 mg asan intravenous infusion, once every four weeks.

The structure of the active compounds identified by code numbers,generic or trade names may be taken from the actual edition of thestandard compendium “The Merck Index” or from databases, e.g. PatentsInternational (e.g. IMS World Publications). The above-mentionedcompounds, which can be used in combination with a compound of thepresent invention, can be prepared and administered as described in theart, such as in the documents cited above.

In one embodiment, the invention provides a product comprising acompound of the present invention and at least one other therapeuticagent as a combined preparation for simultaneous, separate or sequentialuse in therapy. In one embodiment, the therapy is the treatment of adisease or condition mediated by WRN. Products provided as a combinedpreparation include a composition comprising the compound of formula (I)and the other therapeutic agent(s) together in the same pharmaceuticalcomposition, or the compound of the present invention and the othertherapeutic agent(s) in separate form, e.g. in the form of a kit.

In one embodiment, the invention provides a kit comprising two or moreseparate pharmaceutical compositions, at least one of which contains acompound of the present invention. In one embodiment, the kit comprisesmeans for separately retaining said compositions, such as a container,divided bottle, or divided foil packet. An example of such a kit is ablister pack, as typically used for the packaging of tablets, capsulesand the like.

The kit of the invention may be used for administering different dosageforms, for example, oral and parenteral, for administering the separatecompositions at different dosage intervals, or for titrating theseparate compositions against one another. To assist compliance, the kitof the invention typically comprises directions for administration.

In the combination therapies of the invention, the compound of thepresent invention and the other therapeutic agent may be manufacturedand/or formulated by the same or different manufacturers. Moreover, thecompound of the present invention and the other therapeutic may bebrought together into a combination therapy: (i) prior to release of thecombination product to physicians (e.g. in the case of a kit comprisingthe compound of the present invention and the other therapeutic agent);(ii) by the physician themselves (or under the guidance of thephysician) shortly before administration; (iii) in the patientthemselves, e.g. during sequential administration of the compound of thepresent invention and the other therapeutic agent.

Accordingly, the invention provides the use of a compound of the presentinvention for treating a disease or condition mediated by WRN, whereinthe medicament is prepared for administration with another therapeuticagent. The invention also provides the use of another therapeutic agentfor treating a disease or condition mediated by WRN, wherein themedicament is administered with a compound of the present invention.

The invention also provides a compound of the present invention for usein treating a disease or condition mediated by WRN, wherein the compoundof the present invention is prepared for administration with anothertherapeutic agent. The invention also provides another therapeutic agentfor use in treating a disease or condition mediated by WRN, wherein theother therapeutic agent is prepared for administration with a compoundof the present invention. The invention also provides a compound of thepresent invention for use in treating a disease or condition mediated byWRN, wherein the compound of the present invention is administered withanother therapeutic agent. The invention also provides anothertherapeutic agent for use in a method of treating a disease or conditionmediated by WRN, wherein the other therapeutic agent is administeredwith a compound of the present invention.

The invention also provides the use of a compound of the presentinvention for treating a disease or condition mediated by WRN, whereinthe patient has previously (e.g. within 24 hours) been treated withanother therapeutic agent. The invention also provides the use ofanother therapeutic agent for treating a disease or condition mediatedby WRN, wherein the patient has previously (e.g. within 24 hours) beentreated with compound of the present invention.

Example Formulation

A compound of formula (I) may be formulated as an amorphous soliddispersion tablet, as described below.

A. Amorphous Solid Dispersion:

Composition Component Function % (by weight) Example 42 compound (freeform) Active 40 compound Amino methacrylate copolymer Binder 15 (e.g.Eudragit ® E PO)* Copovidone Binder 45 *Eudragit ® E PO is described indx.doi.org/10.1021/mp4000635 I Mol. Pharmaceutics 2013, 10, 2630-2641and has a registry CAS Registry Number of 24938-16-7.

Dichloromethane was charged into a vessel, followed by the compound ofExample 42 as the free form (not the sodium salt), basicpolymethacrylate and copovidone, to obtain a slurry. Dichloromethane wasagain charged to the vessel and the mixture was stirred until a clearsolution was obtained. The solution was spray-dried, then the resultingpowder dried in an agitated bed vacuum drier, then sieved.

B. Film Coated Tablet:

Composition Component Function % (by weight) Example 42 spray driedpowder (from Active 28.98 part A, above) compound Mannitol Filler 59.41Croscarmellose sodium (such as Sodium Disintegrant 5.80 CMC XL Colloidalsilicon dioxide (such as Glidant 0.48 Aerosil ® 200 PH) Sodium stearylfumarate lubricant 1.93 Film-coating agent (such as Opadry ®) Coatingagent 3.4 water

Mannitol, croscarmellose sodium, colloidal silicon dioxide and sodiumstearyl fumarate were added to the Example 42 compound powder obtainedin part A above, and the mixture blended then compressed into tablets.The coating agent was dispersed in water to obtain a homogeneoussuspension, which was used to coat the tablets.

Biological Assays and Data

The activity of a compound according to the present invention can beassessed by the following in vitro & in vivo methods.

Material and Methods

Molecular Biology and virus production. The DNA encoding human Wernerhelicase (UniProt Q14191, WRN, amino acids S2-S1432) was designed asfour DNA strings which were codon-optimized for expression in E. coli.The strings were either ordered from GeneArt (LifeTechnologies,Regensburg, Germany) or made with subcloning overlappingoligonucleotides.

The baculovirus from expression plasmid pLAF1202 (SEQ ID NO: 1) encodingHis-ZZ-3C-WRN (aa N517-P1238, encoded by nucleotides 578-2743 in thesequence) was generated with the FlashBac Ultra system (OxfordExpression Technologies 100302) using 540 ng of plasmid DNA, 5.4 μgFlashbac Ultra DNA, and 5.4 microliters Lipofectin (LifeTechnologies18292-011) for transfection following the manufacturer's instructions.After 5 hours incubation the solution was diluted with 500 microlitersTC100 medium (LifeTechnologies 13055-025) and incubated for 7 days at27° C.

The cells were harvested by centrifugation at 800×g for 10 minutes andthe supernatant containing the virus was transferred into a new steriletube. For the first virus amplification, 500 microliters of the viruswas added to 25 mL of SF9 cells at one million cells/mL and incubatedfor 5 days at 27° C. (200 rpm). The cell viability, density, anddiameter was measured and the virus, upon signs of infection, washarvested by centrifugation at 3000 rpm for 15 minutes.

Baculovirus infected insect cells (BIICs) were generated as described byWasilko et al., 2009, DOI: 10.1016/j.pep.2009.01.002.

In brief, in an Erlenmeyer flask 100 million SF9 cells (one millioncells/mL) in 100 mL ESF921 medium (Expression Systems—96-001-01,supplemented with 0.5× Streptomycin/Penicillin) were infected with 300million baculovirus particles of the respective construct (estimatedMOI=3) and incubated at 27° C. for 24 hours at 130 rpm. The infectedcells were transferred to 50 mL tubes and harvested by centrifugation at100×g for 10 minutes at RT.

The cells were resuspended to 10 million/mL in ESF921 (0.5×Streptomycin/Penicillin) medium with BSA (final 10 mg/mL) and 10% DMSO.500 μL aliquots of cells were transferred to 1.8 mL cryotubes and frozenin Nunc Cryo 1° C. freezing container overnight at −80° C.

SEQ ID NO: 1 AACCATCTCGCAAATAAATAAGTATTTTACTGTTTTCGTAACAGTTTTGTAATAAAAAAACCTATAAATAT TCCGGATTATTCATACCGTCCCACCATCGGGCGCCATGGCTTCTCACCACCATCACCATCACCATCATCA TCACGCTCAGCACGACGAGGCTGTGGACAACAAGTTCAACAAGGAGCAGCAGAACGCTTTCTACGAGATC CTGCACCTCCCTAACCTGAACGAGGAGCAGCGTAACGCTTTCATCCAGTCCCTGAAGGACGACCCTTCTC AGTCTGCTAACCTGCTGGCTGAGGCTAAGAAGCTGAACGACGCTCAGGCTCCTAAGGTGGACAACAAGTT CAACAAGGAGCAGCAGAACGCTTTCTACGAGATCCTGCACCTCCCTAACCTGAACGAGGAGCAGCGTAAC GCTTTCATCCAGTCCCTGAAGGACGACCCTTCTCAGTCTGCTAACCTGCTGGCTGAGGCTAAGAAGCTGA ACGACGCTCAGGCTCCTAAGGTGGACGCTAACGGTGGCGGCGGTTCCGGCGGTGGTGGCTCTCTCGAGGT TCTGTTCCAGGGTCCGAATGAAGGCGAGGAAGATGATGACAAAGACTTCCTGTGGCCAGCTCCAAACGAA GAACAGGTGACTTGTCTCAAGATGTACTTCGGTCATAGCAGCTTCAAACCAGTGCAGTGGAAAGTTATCC ACAGCGTTCTTGAAGAACGTCGTGATAATGTGGCTGTGATGGCTACTGGCTATGGTAAGAGCCTGTGTTT CCAGTACCCGCCAGTTTACGTTGGTAAGATCGGTCTGGTGATTAGCCCGCTGATCTCTCTGATGGAAGAC CAGGTGCTGCAACTTAAGATGAGCAACATCCCGGCTTGTTTCCTGGGTTCTGCACAAAGCGAGAACGTGC TCACCGATATCAAGCTGGGTAAGTACCGTATCGTGTACGTGACGCCAGAATACTGTAGCGGCAACATGGG TCTTCTGCAACAGCTCGAAGCTGATATTGGCATCACCCTCATTGCAGTGGACGAAGCTCACTGTATCAGC GAGTGGGGTCATGATTTCCGCGACTCTTTCCGTAAACTGGGTTCTCTGAAGACTGCACTTCCGATGGTTC CAATTGTGGCACTGACCGCAACTGCTTCTAGCTCTATTCGTGAAGACATCGTTCGTTGCCTGAACCTCCG TAACCCACAAATTACCTGCACCGGCTTTGACCGTCCGAACCTGTACCTGGAGGTTCGTCGTAAGACCGGT AATATCCTTCAGGACCTGCAACCATTCCTGGTTAAGACCAGCAGCCACTGGGAGTTCGAAGGTCCGACTA TCATCTACTGCCCAAGCCGTAAGATGACCCAGCAGGTTACTGGTGAACTGCGTAAACTGAACCTGAGCTG TGGCACTTACCACGCAGGCATGTCTTTCTCTACCCGTAAAGACATCCATCATCGTTTCGTGCGTGATGAA ATCCAGTGCGTTATCGCTACCATTGCATTCGGCATGGGTATCAACAAAGCTGACATCCGTCAAGTGATTC ACTACGGTGCACCGAAAGACATGGAAAGCTACTACCAGGAAATCGGCCGTGCAGGTCGTGATGGTCTGCA AAGCTCTTGTCATGTGCTGTGGGCACCAGCAGATATTAACCTGAACCGTCACCTGCTGACTGAAATTCGT AACGAGAAATTCCGTCTGTACAAACTGAAGATGATGGCGAAGATGGAGAAATACCTGCATAGCTCCCGTT GTCGTCGTCAAATCATTCTGAGCCATTTCGAGGATAAACAGGTGCAGAAAGCTTCTCTGGGTATCATGGG CACTGAGAAGTGCTGCGATAACTGTCGTAGCCGTCTTGATCACTGCTACAGCATGGACGATAGCGAAGAC ACTTCTTGGGATTTCGGTCCACAAGCATTCAAACTGCTGAGCGCAGTTGATATCCTGGGTGAGAAATTCG GCATCGGCCTCCCAATCCTGTTTCTGCGCGGTTCTAACTCTCAGCGTCTTGCTGATCAATACCGTCGTCA CTCTCTGTTCGGCACTGGTAAAGACCAGACCGAATCTTGGTGGAAAGCATTCAGCCGTCAACTTATCACC GAAGGCTTTCTGGTGGAAGTGTCTCGTTACAACAAGTTCATGAAGATCTGCGCACTGACTAAGAAAGGTC GTAACTGGCTGCACAAGGCAAATACCGAGTCTCAGTCTCTTATCCTTCAGGCTAACGAAGAACTGTGCCC GAAGAAGCTTCTGCTGCCATCTTCTAAGACCGTGAGCTCTGGTACTAAAGAGCATTGCTACAACCAGGTG CCGGTTGAACTGTCTACCGAGAAGAAGTCCAACCTGGAGAAGCTGTACTCCTACAAACCGTGCGACAAGA TCTCCTCCGGTTCTAATATCAGCAAGAAGTCCATCATGGTGCAGTCTCCGGAGAAAGCTTACAGCAGCTC TCAGCCAGTTATCTCTGCACAGGAACAGGAAACTCAGATTGTGCTGTACGGTAAACTGGTGGAAGCACGT CAGAAACACGCTAACAAGATGGACGTGCCGCCAGCAATTCTTGCAACCAACAAGATTCTGGTGGACATGG CTAAGATGCGCCCAACTACTGTTGAGAACGTGAAACGTATCGACGGTGTTAGCGAAGGTAAAGCTGCAAT GCTGGCACCACTGCTTGAAGTTATCAAGCATTTCTGCCAGACCAACTCTGTTCAGACCGACCTGTTCTCT TCTACCAAACCATAATGGTACCGAATTCGCGGCCGCAGAGCTCGCTCTGGTGCCACGCGGTAGTTCCGCT TGGAGCCACCCGCAGTTCGAAAAGTAAGTGATTAACCTCAGGTTATACATATATTTTGAATTTAATTAAT TATACATATATTTTATATTATTTTTGTCTTTTATTATCGAGGGGCCGTTGTTGGTGTGGGGTTTTGCATA GAAATAACAATGGGAGTTGGCGACGTTGCTGCGCCAACACCACCTCCCTTCCCTCCTTTCATCATGTATC TGTAGATAAAATAAAATATTAAACCTAAAAACAAGACCGCGCCTATCAACAAAATGATAGGCATTAACTT GCCGCTGACGCTGTCACTAACGTTGGACGATTTGCCGACTAAACCTTCATCGCCCAGTAACCAATCTAGG TAGCTGAGCGCATGCAAGCTGATCCGGGTTATTAGTACATTTATTAAGCGCTAGATTCTGTGCGTTGTTG ATTTACAGACAATTGTTGTACGTATTTTAATAATTCATTAAATTTATAATCTTTAGGGTGGTATGTTAGA GCGAAAATCAAATGATTTTCAGCGTCTTTATATCTGAATTTAAATATTAAATCCTCAATAGATTTGTAAA ATAGGTTTCGATTAGTTTCAAACAAGGGTTGTTTTTCCGAACCGATGGCTGGACTATCTAATGGATTTTC GCTCAACGCCACAAAACTTGCCAAATCTTGTAGCAGCAATCTAGCTTTGTCGATATTCGTTTGTGTTTTG TTTTGTAATAAAGGTTCGACGTCGTTCAAAATATTATGCGCTTTTGTATTTCTTTCATCACTGTCGTTAG TGTACAATTGACTCGACGTAAACACGTTAAATAGAGCTTGGACATATTTAACATCGGGCGTGTTAGCTTT ATTAGGCCGATTATCGTCGTCGTCCCAACCCTCGTCGTTAGAAGTTGCTTCCGAAGACGATTTTGCCATA GCCACACGACGCCTATTAATTGTGTCGGCTAACACGTCCGCGATCAAATTTGTAGTTGAGCTTTTTGGAA TTATTTCTGATTGCGGGCGTTTTTGGGCGGGTTTCAATCTAACTGTGCCCGATTTTAATTCAGACAACAC GTTAGAAAGCGATGGTGCAGGCGGTGGTAACATTTCAGACGGCAAATCTACTAATGGCGGCGGTGGTGGA GCTGATGATAAATCTACCATCGGTGGAGGCGCAGGCGGGGCTGGCGGCGGAGGCGGAGGCGGAGGTGGTG GCGGTGATGCAGACGGCGGTTTAGGCTCAAATGTCTCTTTAGGCAACACAGTCGGCACCTCAACTATTGT ACTGGTTTCGGGCGCCGTTTTTGGTTTGACCGGTCTAAGACGAGTGCGATTTTTTTCGTTTCTAATAGCT TCCAACAATTGTTGTCTGTCGTCTAAAGGTGCAGCGGGTTGAGGTTCCGTCGGCATTGGTGGAGCGGGCG GCAATTCAGACATCGATGGTGGTGGTGGTGGTGGAGGCGCTGGAATGTTAGGCACGGGAGAAGGTGGTGG CGGCGGTGCCGCCGGTATAATTTGTTCTGGTTTAGTTTGTTCGCGCACGATTGTGGGCACCGGCGCAGGC GCCGCTGGCTGCACAACGGAAGGTCGTCTGCTTCGAGGCAGCGCTTGGGGTGGTGGCAATTCAATATTAT AATTGGAATACAAATCGTAAAAATCTGCTATAAGCATTGTAATTTCGCTATCGTTTACCGTGCCGATATT TAACAACCGCTCAATGTAAGCAATTGTATTGTAAAGAGATTGTCTCAAGCTCGGATCGATCCCGCACGCC GATAACAAGCCTTTTCATTTTTACTACAGCATTGTAGTGGCGAGACACTTCGCTGTCGTCGAGGTTTAAA CGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAG GCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAA AGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCA CAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCT GGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTT CGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAA GCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAG TCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGT ATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGG TATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACC ACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAG ATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCAT GAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGT ATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTC TATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATC TGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAG CCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTT GCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCAT CGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACA TGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGG CCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATG CTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCT TGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAAC GTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGC ACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAAT GCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATT GAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAAT AGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGCGCCCTGTAGCGGCGCATTAAGCGCGGCG GGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCT TCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTT CCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCA TCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCC AAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGC CTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACA ATTTCCCATTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTAC GCCAGGAACGGCTCCGCCCACTATTAATGAAATTAAAAATTCCAATTTTAAAAAACGCAGCAAGAGAAAC ATTTGTATGAAAGAATGCGTAGAAGGAAAGAAAAATGTCGTCGACATGCTGAACAACAAGATTAATATGC CTCCGTGTATAAAAAAAATATTGAACGATTTGAAAGAAAACAATGTACCGCGCGGCGGTATGTACAGGAA GAGGTTTATACTAAACTGTTACATTGCAAACGTGGTTTCGTGTGCCAAGTGTGAAAACCGATGTTTAATC AAGGCTCTGACGCATTTCTACAACCACGACTCCAAGTGTGTGGGTGAAGTCATGCATCTTTTAATCAAAT CCCAAGATGTGTATAAACCACCAAACTGCCAAAAAATGAAAACTGTCGACAAGCTCTGTCCGTTTGCTGG CAACTGCAAGGGTCTCAATCCTATTTGTAATTATTGAATAATAAAACAATTATAAATGCTAAATTTGTTT TTTATTAACGATACAAACCAAACGCAACAAGAACATTTGTAGTATTATCTATAATTGAAAACGCGTAGTT ATAATCGCTGAGGTAATATTTAAAATCATTTTCAAATGATTCACAGTTAATTTGCGACAATATAATTTTA TTTTCACATAAACTAGACGCCTTGTCGTCTTCTTCTTCGTATTCCTTCTCTTTTTCATTTTTCTCTTCAT AAAAATTAACATAGTTATTATCGTATCCATATATGTATCTATCGTATAGAGTAAATTTTTTGTTGTCATA AATATATATGTCTTTTTTAATGGGGTGTATAGTACCGCTGCGCATAGTTTTTCTGTAATTTACAACAGTG CTATTTTCTGGTAGTTCTTCGGAGTGTGTTGCTTTAATTATTAAATTTATATAATCAATGAATTTGGGAT CGTCGGTTTTGTACAATATGTTGCCGGCATAGTACGCAGCTTCTTCTAGTTCAATTACACCATTTTTTAG CAGCACCGGATTAACATAACTTTCCAAAATGTTGTACGAACCGTTAAACAAAAACAGTTCACCTCCCTTT TCTATACTATTGTCTGCGAGCAGTTGTTTGTTGTTAAAAATAACAGCCATTGTAATAAGACGCACAAACT AATATCACAAACTGGAAATGTCTATCAATATATAGTTGCTGATCAGATCTACCCGTAGTGGCTATGGCAG GGCTTGCCGCCCCGACGTTGGCTGCGAGCCCTGGGCCTTCACCCGAACTTGGGGGTTGGGGTGGGGAAAA GGAAGAAACGCGGGCGTATTGGTCCCAATGGGGTCTCGGTGGGGTATCGACAGAGTGCCAGCCCTGGGAC CGAACCCCGCGTTTATGAACAAACGACCCAACACCCGTGCGTTTTATTCTGTCTTTTTATTGCCGTCATA GCGCGGGTTCCTTCCGGTATTGTCTCCTTCCGTGTTTCAGTTAGCCTCCCCCATCTCCCGGTACCGCATG CTATGCATCGGCCGCTTTACTTGTACAGCTCGTCCATGCCGAGAGTGATCCCGGCGGCGGTCACGAACTC CAGCAGGACCATGTGATCGCGCTTCTCGTTGGGGTCTTTGCTCAGGGCGGACTGGGTGCTCAGGTAGTGG TTGTCGGGCAGCAGCACGGGGCCGTCGCCGATGGGGGTGTTCTGCTGGTAGTGGTCGGCGAGCTGCACGC TGCCGTCCTCGATGTTGTGGCGGATCTTGAAGTTCACCTTGATGCCGTTCTTCTGCTTGTCGGCCATGAT ATAGACGTTGTGGCTGTTGTAGTTGTACTCCAGCTTGTGCCCCAGGATGTTGCCGTCCTCCTTGAAGTCG ATGCCCTTCAGCTCGATGCGGTTCACCAGGGTGTCGCCCTCGAACTTCACCTCGGCGCGGGTCTTGTAGT TGCCGTCGTCCTTGAAGAAGATGGTGCGCTCCTGGACGTAGCCTTCGGGCATGGCGGACTTGAAGAAGTC GTGCTGCTTCATGTGGTCGGGGTAGCGGCTGAAGCACTGCACGCCGTAGGTCAGGGTGGTCACGAGGGTG GGCCAGGGCACGGGCAGCTTGCCGGTGGTGCAGATGAACTTCAGGGTCAGCTTGCCGTAGGTGGCATCGC CCTCGCCCTCGCCGGACACGCTGAACTTGTGGCCGTTTACGTCGCCGTCCAGCTCGACCAGGATGGGCAC CACCCCGGTGAACAGCTCCTCGCCCTTGCTCACCATCGTCGAGATCCCGGGCGTTTAAATTGTGTAATTT ATGTAGCTGTAATTTTTACCTTATTAATATTTTTTACGCTTTGCATTCGACGACTGAACTCCCAAATATA TGTTTAACTCGTCTTGGTCGTTTGAATTTTTGTTGCTGTGTTTCCTAATATTTTCCATCACCTTAAATAT GTTATTGTAATCCTCAATGTTGAACTTGCAATTGGACACGGCATAGTTTTCCATAGTCGTGTAAAACATG GTATTGGCTGCATTGTAATACATCCGACTGAGCGGGTACGGATCTATGTGTTTGAGCAGCCTGTTCAAAA ACTCTGCATCGTCGCAAAACGGAATTTGGTACCCGGGCGTATACTCCGGAATATTAATAGATCATGGAGA TAATTAAAATGAT

Protein Expression and Purification

BIICs aliquots for Werner helicase protein His-ZZ-3C-WRN (aa N517-P1238,pLAF1202) were diluted 1/100 into ESF921 medium and further diluted1/100 into the expression/production flasks with Sf21 cells (one millioncells/mL) in 1 L ESF921 medium and incubated for protein expression for96 h (27° C., 130 rpm).

The WRN protein was purified using the following protocol. The cellpellets were thawed and resuspended in 80 mL buffer A (50 mM Tris, 300mM NaCl, 20 mM imidazole, 1 mM TCEP, 10% glycerol, pH 7.8) supplementedwith Turbonuclease (final concentration 40 units/mL, Merck) and cOmpleteprotease inhibitor tablets (1 tablet/50 mL, Roche). The cells were lysedby three passages through a homogenizer (Avestin, Emulsiflex C3) at800-1000 bar. The lysed sample was centrifuged at 48000×g for 40 minutes(Sorvall RC5B, SS-34 rotor) and the supernatant was passed through a0.45 μm filter.

The lysate was loaded onto a HisTrap crude FF 5 mL column (GEHealthcare) mounted on an ÄKTA Pure 25 chromatography system (GEHealthcare). Contaminating proteins were washed away with buffer A andbound protein was eluted with a linear gradient over 10 column volumesto 100% of buffer B (50 mM Tris, 300 mM NaCl, 300 mM imidazole, 1 mMTCEP, 10% glycerol, pH 7.8). 1% (w/w) HRV 3C protease (His-MBP-tagged,produced in-house) was added to the eluted protein. The N-terminalpurification tag was cleaved off by the protease during dialysisovernight at 5° C. against 2 L buffer (50 mM Tris pH 7.0, 150 mM NaCl, 1mM TCEP, 10% glycerol, 0.02% CHAPS). The protein solution was thencarefully diluted with adding two volume parts of 20 mM Tris pH 7.0, 10%glycerol, 0.02% CHAPS. The slightly turbid protein solution was passedover a 0.45 μm filter. The cleaved protein was loaded onto a Resource S6 mL column (GE Healthcare) pre-equilibrated with 20 mM Tris, 20 mMNaCl, 1 mM TCEP, 10% glycerol, pH 7.0. Cleaved tag and contaminatingproteins were washed away with the equilibration buffer. The boundtarget protein was eluted with a linear gradient over 20 column volumesof the same buffer containing 1 M sodium chloride and then injected ontoa HiLoad 16/600 Superdex 75 pg column (GE Healthcare) pre-equilibratedwith 50 mM Tris pH 7.4, 300 mM NaCl, 10% glycerol. Fractions containingpure protein were identified by SDS-PAGE and pooled. The purifiedprotein was finally split into aliquots and frozen on dry ice. Thepurity, quantity, and identity of the protein was determined by RP-HPLCand LC-MS.

In Vitro Enzymatic Activity Assay on WRN Helicase

An ATPase assay was set up to measure the DNA dependent ATP hydrolysisactivity of WRN helicase. This assay was used also to assess theinhibition properties of compounds of the invention on DNA dependent WRNATPase activity.

The core helicase motif of the WRN protein (aa N517-P1238) was producedfor this assay (protein production as described above). A 45oligonucleotide sequence called “FLAP26” as described by Brosh et al.,2009, DOI: 10.1074/jbc.M111446200(TTTTTTTTTTTTTTTTTTTTTTCCAAGTAAAACGACGGCCAGTGC; SEQ ID NO: 2) waspurchased from IDT (Integrated DNA Technologies, Leuven, Belgium) andused as single strand DNA substrate. The ADP-Glo assay kit (Promega,Madison, Wis.) allowing the quantification of ADP produced in ATPhydrolysis reactions was used for setting up this assay.

Time course experiments were first performed in order to determine thebest enzymatic assay conditions (including buffer conditions, reactiontime and concentrations of protein, ATP and DNA substrates). A typicalreaction consists of 10 nM WRN protein, 0.2 nM FLAP26, and 300micromolar ATP in the following assay buffer: 30 mM Tris pH7.5, 2 mMMgCl₂, 0.02% BSA, 50 mM NaCl, 0.1% pluronic F127 prepared in DNAse freewater. To evaluate the inhibition properties of compounds of theinvention, serial dilutions were prepared in DMSO (10 half log dilutionsfrom a 10 mM DMSO solution). 50 nanoliters of each concentration waspre-incubated for 3 hours in a 384 small volume assay plate (Greiner#784075) with 2.5 microliters of a 20 nM WRN helicase protein in assaybuffer with 600 micromolar ATP. Control wells were included with a “highcontrol” (no inhibition), containing DMSO with no test compound, and“low controls” (maximal inhibition), containing buffer without protein.The reaction was started by addition of 2.5 microliters of FLAP26 at 0.4nM and incubated for 30 minutes at room temperature. The reaction wasstopped with the addition of 5 microliters of the first ADP-Glo reagentand incubated for one hour to remove the excess amount of ATP.Afterwards, 10 microliters of ATP detection reagent was added andincubated for an additional hour before reading. Luminescence output wasrecorded using Tecan 1000 reader, with 5 minutes delay before reading.Each concentration of compound was tested in duplicates in the assayplate.

Data analysis was carried out using an in-house developed software(Novartis Helios software application, Novartis Institutes forBioMedical Research, unpublished) using the methods described byFormenko et al., 2006, DOI: 10.1016/j.cmpb.2006.01.008. Followingnormalization of activity values for the wells to % inhibition (%inhibition=[(high control-sample)/(high control-low control)]×100), IC₅₀fitting was carried out from the duplicate determinations present oneach plate according to [4]. Data analysis can also be carried out usingcommercially available software designed to derive IC₅₀ values using4-parameter fits (e.g. GraphPad Prism, XL fit). The reported IC₅₀ valuesare the geometrical means of at least 2 independent replicates.

Method for Detecting Effects on Cellular Proliferation

The colon carcinoma cell lines SW48 (RRID: CVCL_1724), HCT 116 (RRID:CVCL_0291) and SNU-407 (RRID: CVCL_5058) were obtained from ATCC. TheWRN-knockdown insensitive colon carcinoma cell line DLD-1 (RRID:CVCL_0248) was obtained from the Korean Cell Line Bank (KCLB), and usedto generate a derivative in which the endogenous WRN gene copies wereknocked out by CRISPR-mediated editing using standard CRISPR methods.The resulting cell line, DLD1-WRN—KO, was used to assess potentialoff-target compound effects.

SW48, SNU-407 and DLD1-WRN—KO cells were cultured in growth mediumcomposed of RPMI-1640 (Amimed Cat #1-41F22-I), 2 mM L-Glutamine (AmimedCat #5-10K50), 10 mM HEPES (Gibco Cat #15630-056), 1 mM sodium pyruvate(Amimed Cat #5-60F00-H), 1× Penicillin-Streptomycin (Amimed Cat #4-01F00-H) and 10% fetal calf serum (Amimed Cat #2-01 F30-G, Lot #LB11566P).HCT 116 cells were cultured in growth medium composed of McCoys 5A(Amimed catalog #1-18F01-I), 2 mM L-Glutamine (Amimed Cat #5-10K50), 1×Penicillin-Streptomycin (Amimed Cat #4-01 F00-H) and 10% fetal calfserum (Amimed Cat #2-01F30-G, Lot #LB11566P). All cells were maintainedat 37° C. in a humidified 5% CO₂ incubator.

Following filtration through a Steriflip-NY 20 μm filter (Millipore Cat# SCNY00020), trypsinized cells were seeded in 100 microliters growthmedium at 2′000 (SW48) or 1500 (SNU-407, DLD1-WRN—KO, HCT 116)cells/well into white, clear-bottom 96-well plates (Costar Cat #3903).Three replicate plates were prepared for each compound treatmentcondition. In addition, one plate (termed “day 0”) was prepared toquantify the number of viable cells at the time of compound addition.Following overnight incubation at 37° C. in a humidified 5% CO₂atmosphere, eight 3-fold serial dilutions of a given compound stock(obtained at a concentration of 10 mM in DMSO and stored at 4° C.) weredispensed directly into each of the triplicate assay plates using a HP300D non-contact Digital Dispenser (TECAN). The final concentration ofDMSO was normalized to 0.1% in all wells. 96 hours after compoundaddition, cellular ATP levels as a surrogate for cell viability wasassessed following addition of 50 microliters CellTiterGlo (Promega Cat#G7573) reagent and luminescence quantification on a MPLEX multi-modeplate-reader (TECAN) following a 10 minute incubation at roomtemperature. The number of viable cells in the “day 0” plate werequantified identically on the day of compound addition.

For data analysis, the assay background signal that was determined inwells containing medium, but no cells, was subtracted from all otherdata points prior to further calculations. The extent of growthinhibition and potential cell kill was assessed by comparing the ATPlevels (measured using CellTiterGlo, Promega) in compound-treated cellswith those present at the time of compound addition. To this end, thefollowing conditional concept was programmatically applied in HELIOS, anin-house software applying a multi-step decision tree to arrive atoptimal concentration response curve fits (Gubler et al, SLAS DOI:10.1177/2472555217752140) to calculate % growth (% G) for eachcompound-treated well: % G=(T−V0)/V0))*100 when T<V0, and %G=(T−V0)/(V−V0)))*100 when T≥V0, where V0 is the viability level at timeof compound addition, while V and T represent vehicle-control andcompound-treated viability levels, respectively, at the end of thecompound incubation. 100%, 0% and −100% signify absence of growthinhibition, growth stasis, and complete cell kill, respectively.Compound concentrations leading to half-maximal growth inhibition (G150)and residual cell viability at the highest tested compound concentration(Data (cmax), expressed in percent) were routinely calculated. Dataanalysis can also be carried out using commercially available softwaredesigned to derive IC50 values using 4-parameter fits (e.g. GraphPadPrism, XL fit). The reported GI₅₀ values are the geometrical means of atleast 2 independent replicates.

In Vivo Efficacy Demonstration for Compounds of the Invention

Experiments were performed in female Crl:NU(NCr)-Foxn1nu-Homozygous nudemice (Charles River). Animals were housed under Optimized HygienicConditions in Allentown XJ cages (IVC, max. 6 mice per cage) with foodand water at libitum and a 12 h:12 h light: dark cycle. Animals wereallowed to acclimatize for at least 1 week before being enrolled in theexperimental design. The study described here was performed according tolicense 2275 approved by the Basel Cantonal Veterinary Office.

SW48 human colorectal cancer cells were obtained from ATCC. The cellswere cultured in RPMI-1640 medium (BioConcept Ltd. Amimed, #1-41F01-I)supplemented with 10% FCS (Bio Concept #2-01 F30-I), 2 mM L-glutamine(BioConcept Ltd. Amimed, #5-10K50-H), 1 mM sodium pyruvate (Bio Concept#5-60F00-H) and 10 mM HEPES (Bio Concept #5-31 F00-H) at 37° C. in anatmosphere of 5% CO₂ in air. To establish SW48 xenografts cells wereharvested and re-suspended in HBSS (Gibco, #14175) before injecting 100μL containing 5 million cells subcutaneously in the right flank ofanimals which were anesthetized with isoflurane.

Tumour growth was monitored regularly post cell inoculation and animalswere randomised into treatment groups (n=7) when tumor volume reachedappropriate volume. During the treatment period, tumor volume wasmeasured about twice a week. Tumor size, in mm³, was calculated from:(L×W2×π/6). Where W=width and L=length of the tumor.

Depending on the target concentration, 50-200 mg of amorphous sodiumsalt of test compound (corrected by salt factor) were dissolved in 8 mLof an aqueous 10% w/v solution of 2-hydroxypropyl-beta-cyclodextrin(HPBCD). The pH was adjusted to pH 7.4 with 0.1 M HCl (˜1 eq.) and theresulting solution filled up to a total volume of 10 mL with aqueous 10%HPBCD solution. In case of turbidity, the solution was filtered. Theresulting solution formulation was used for in vivo studies.

Tumor bearing animals were enrolled into treatment groups (n=7) whentheir tumors reached an appropriate size to form groups with a meantumor volume of 186 mm³.

Animals were then treated with vehicle (10%Hydroxypropyl-beta-Cyclodextrine) or 240 mg/kg of a compound of theinvention daily (QD) by oral gavage at 20 mL/kg.

Animals were weighed twice per week and examined frequently for overtsigns of any adverse effects.

Tumor and body weight change data were analyzed statistically usingGraphPad Prism 7.00 (GraphPad Software). If the variances in the datawere normally distributed, the data were analyzed using one-way ANOVAwith post hoc Dunnett's test for comparison of treatment versus controlgroup. When applicable, results are presented as mean±SEM.

As a measure of efficacy the % T/C value is calculated at the end of theexperiment according to:

(Δtumor volume^(treated)/Δtumor volume^(control))*100

Tumor regression was calculated according to:

−(Δtumor volume^(treated)/tumor volume^(treated at start))*100

Where Δtumor volumes represent the mean tumor volume on the evaluationday minus the mean tumor volume at the start of the experiment.

Treatment was initiated with, for example, Example 58 in 10%Hydroxypropyl-beta-Cyclodextrine in water at 240 mg/kg using QD oralapplication when the average tumor volume was 186 mm³ (n=7/group). Thetreatment period with Example 58 was 18 days after which overallefficacy and tolerability were evaluated based on tumor volume and bodyweight changes observed during the treatment period (FIG. 6 ). Example58 dosed orally at 240 mg/kg qd induced an antitumor response againstSW48 xenografts in nude mice (FIG. 6 ). The T/C % value on day 18 was −9(p=<0.05 when compared with vehicle control, one way ANOVA withDunnett's post hoc test). The mean tumor volume on day 18 in the 4/7surviving animals showed a −26% regression. Based on body weight, QDdosing of 240 mg/kg of Example 58 was well tolerated (FIG. 6 ). Data forthe compounds of Examples 42, 96 and 57 are presented in FIGS. 9, 10 and11 respectively.

The instant application contains Sequence Listings which have beensubmitted electronically in ASCII format and are hereby incorporated byreference in their entirety. Said ASCII copy, created on 21 May 2021, isnamed PAT059096_SL.txt and is 13184 bytes in size.

The following table shows the IC₅ data in the WRN ATPase assay and theGI₅₀ data for the proliferation assays using SW48 and DLD1-WRN—KO celllines for compounds of the invention. For example, Example 1 is a 50 nMWRN ATPase inhibitor with a proliferation GI₅₀ of 50 nM in the SW4and >10 micromolar in the DLD1 WRN—KO cell lines.

Proliferation Proliferation Proliferation WRN Proliferation Assay WRNAssay Assay ATPase Assay DLD1- ATPase SW48 DLD1- Activity SW48 WRN-KOActivity (GI₅₀) WRN-KO (IC₅₀) (GI₅₀) (GI₅₀) Ex. (IC₅₀) μM μM (GI₅₀) μMEx. μM μM μM  1 0.05 0.05 >10  61 0.05 0.05 >10  2 0.08 0.19 >10  620.05 0.05 >10  3 0.07 0.09 >10  63 0.11 0.05 >10  4 0.04 0.06 >10  640.09 0.48 >10  5 0.09 0.12 >10  65 0.05 0.08  7.8  6 0.07 0.27 >10  660.04 0.09 >10  7 0.09 0.21 >10  67 0.15 0.19 >10  8 0.08 0.21 >10  680.12 0.27 >10  9 0.06 0.35 >10  69 0.04 0.03  6.3  10 0.25 0.29  9.9  700.10 0.22 >10  11 0.11 0.38 >10  71 0.07 0.06 >10  12 0.13 0.55 >10  720.05 0.05 >10  13 0.03 0.16  5.2  73 0.07 0.20 >10  14 0.14 0.48 >10  740.05 0.07 >10  14a 0.08 0.37 >10  75 0.05 0.08 >10  14b 0.09 0.26 >10 15 0.11 0.44 >10  76 0.08 0.19 >10  15a 0.14 0.40 >10  77 0.21 0.38 >10 15b 0.12 0.36 >10  78 0.08 0.30 >10  16 0.23 0.32 >10  79 0.04 0.50 >10 16a 0.19 0.34 >10  80 0.06 0.07 >10  16b 0.18 0.23 >10  80a 0.070.07 >10  17 0.14 0.24 >10  80b 0.08 0.06 >10  18 0.18 0.26 >10  81a0.04 0.07 >10  19 0.09 0.11  5  81b 0.04 0.06 >10  20 0.31 0.10 >10  820.28 7.72 >10  21 0.18 1.07 >10  83 0.09 0.06 >10  22 0.60 2.63 >10  840.05 0.13 >10  23 0.26 1.54 >10  85 0.07 0.11 >10  24a 0.11 0.04 >10  860.09 0.08 >10  24b 0.10 0.05 >10  87 0.11 0.06 >10  25a 0.16 0.05 >10 88 0.37 0.76 >10  25b 0.10 0.05 >10  89 0.04 0.17 >10  26 0.25 0.08 >10 90 0.13 0.30 >10  27 0.13 0.27 >10  91 0.08 0.32 >10  28 0.13 0.10 >10 92 0.05 0.08 >10  29 0.09 0.05 >10  93 0.08 0.07 >10  30 0.14 0.16 >10 94 0.04 0.05 >10  31 0.23 0.32 >10  95 0.05 0.06 >10  32 0.06 0.06  7.6 96 0.06 0.06 >10  33 0.26 0.22 >10  97 0.08 0.07 >10  34 0.25 0.28 >10 98 0.04 0.14 >10  35 0.19 0.40 >10  99 0.18 0.28 >10  36 0.93 1.78 >10100 0.06 0.03 >10  37 0.02 0.03 >10 101 0.10 0.21 >10  38 0.04 0.05 >10102 0.09 0.17 >10  39 0.04 0.05 >10 103 0.10 0.25 >10  40 0.02 0.03 >10104 0.07 0.05 >10  41 0.04 0.04 >10 105 0.20 1.13 >10  42 0.10 0.05 >10106 0.06 0.11 >10  43 0.07 0.05 >10 107 0.05 0.10 >10  44 0.11 0.08 >10108 0.08 0.09 >10  45 0.13 0.08 >10 109 1.18 0.69 >10  46 0.11 0.11 >10110 0.36 1.21 >10  47 0.14 0.17 >10 111 0.33 1.12 >10  48 0.07 0.08 >10112 0.51 7.94 >10  49 0.12 0.12 >10 113 0.04 0.12 >10  50 0.03 0.05 >10114 0.09 0.18 >10  51 0.04 0.03 >10 115 0.17 0.20 >10  52 0.04 0.04 >10116 0.06 0.08  6.6  53 0.12 0.14 >10 117 0.04 0.26 >10  54 1.30 6.01 >10118 0.09 0.45 >10  55 0.20 0.10 >10 119 0.16 0.05 >10  56 0.16 0.25 >10120 0.47 0.93 >10  57 0.10 0.10 >10 121 0.86 1.74 >10  58 0.06 0.07 >10122 0.15 0.10 >10  59 0.80 123 0.10 0.05 >10  60 0.24 0.88 >10 124 0.360.39 >10 126 0.13 125 0.03 0.01 >10 127 0.08 0.09 >10 134 0.06 0.21 >10128 0.08 0.69 >10 135 0.13 0.28 >10 129 0.50 2.55 >10 136 0.08 0.65 >10130 0.10 0.05 >10 137 0.05 0.24  6.1 131 0.11 0.03 >10 132 0.89 0.49 >10133 0.12 0.06 >10

Data are geometric means with at least 2 duplicate determinations.

In another aspect, the invention provides a compound of formula (I), foruse in the treatment of cancer, or as a research chemical such as achemical probe, wherein part of the WRN inhibition activity is via amechanism which is not assessed by the biochemical assay describedabove.

Method for Detecting Clonogenic Effects on Cellular Proliferation(CFA=Colony Formation Assay)

Cell lines were obtained from ATCC and media and culture conditions usedas recommended by ATCC. All cells were maintained at 37° C. in ahumidified 5% CO₂ incubator. Cells were seeded at 250-2,000 cells perwell in a 12-well plate in 1 ml of media. The WRN inhibitor compound ofexample 42 was added with a starting concentration of 10 uM. Followingovernight incubation at 37° C. in a humidified 5% CO₂ atmosphere, ten3-fold serial dilutions of a given compound stock (obtained at aconcentration of 10 mM in DMSO and stored at 4° C.) were dispenseddirectly into each assay plates using a HP 300D non-contact DigitalDispenser (TECAN). The final concentration of DMSO was normalized to0.1% in all wells. Cells were left in the incubator for 8-20 days, withmedium exchange every 3-4 days. After this time, 100 μl formaldehyde 37%was added directly in each test well and incubated for 15 minutes atroom temperature. After rinsing twice with 5 ml of water, 0.5 ml of0.05% methylene blue was added for 20 minutes at room temperature. Wellswere rinsed three times with water and 1 ml of 3% HCl added to theplates and shaken until complete color dissolution. 200 μl of thissolution was transferred in a 96 well plate and absorbance measured at650 nM using microtiter plate reader (Synergy HT). Compoundconcentrations leading to half-maximal growth inhibition (G150) werecalculated using XLfit using the Dose Response One Site model 201, withfit=(A+((B−A)/(1+((x/C){circumflex over ( )}D)))). For non-adherent celllines, cellular ATP levels as a surrogate for cell viability wereassessed following addition of 200 μl of CellTiterGlo (Promega Cat#G7573) reagent to the culture after removal of 600 μl. Luminescencequantification was performed on a Synergy HT plate-reader following a 15min incubation at room temperature. Data were analyzed as for themethylene blue stain. Results are shown in FIG. 8 . Further informationregarding MSI-H and MSS is available in the following references:

-   -   Chan et Al., WRN helicase is a synthetic lethal target in        microsatellite unstable cancers. Nature. 2019 April;        568(7753):551-556. doi: 10.1038/s41586-019-1102-x. Epub 2019        Apr. 10. PMID: 30971823; PMCID: PMC6580861.    -   McDonald E. R. et al., Project DRIVE: A Compendium of Cancer        Dependencies and Synthetic Lethal Relationships Uncovered by        Large-Scale, Deep RNAi Screening. Cell 170(3):577-592 (2017)).

Preparation of Compounds

Compounds of the present invention can be prepared as described in thefollowing Examples. The Examples are intended to illustrate theinvention and are not to be construed as being limitations thereof.

Instrumentation

Microwave: All microwave reactions were conducted in a Biotage Initiatoror an Anton Paar monowave 450, irradiating at 0-400 W from a magnetronat 2.45 GHz with Robot Eight/Robot Sixty/Robot twenty four processingcapacity, unless otherwise stated.

UPLC-MS Methods: Using Waters Acquity UPLC with Waters SQ detector,unless stated otherwise.

UPLC-MS 1:

Column CORTECS ™ C18+ 2.7 μm, Column Dimension 2.1 × 50 mm ColumnTemperature 80° C. Eluents A: water + 4.76% isopropanol + 0.05% FA +3.75 mM AA B: isopropanol + 0.05% FA Flow Rate 1.0 mL/min Gradient 1 to50% B in 1.4 min; 50 to 98% B in 0.3 min

UPLC-MS 2:

Column ACQUITY UPLC ® BEH C18 1.7 μm Column Dimension 2.1 × 100 mmColumn Temperature 80° C. Eluents A: water + 4.76% isopropanol + 0.05%FA + 3.75 mM AA B: isopropanol + 0.05% FA Flow Rate 0.4 mL/min Gradient1 to 60% B in 8.4 min; 60 to 98% B in 1.0 min

UPLC-MS 3:

Column ACQUITY UPLC ® BEH C18 1.7 μm Column Dimension 2.1 × 50 mm ColumnTemperature 80° C. Eluents A: water + 4.76% isopropanol + 0.05% FA +3.75 mM AA B: isopropanol + 0.05% FA Flow Rate 0.6 mL/min Gradient 1 to98% B in 1.7 min

UPLC-MS 4:

Column ACQUITY UPLC ® BEH C18 1.7 μm Column Dimension 2.1 × 50 mm ColumnTemperature 80° C. Eluents A: water + 0.05% FA + 3.75 mM AA B:isopropanol + 0.05% FA Flow Rate 0.6/0.7 mL/min Gradient 5 to 98% B in1.7 min

UPLC-MS 5:

Column XBridge ® BEH ™ C18 2.5 μm Column Dimension 2.1 × 50 mm ColumnTemperature 80° C. Eluents A: water + 5 mM NH₄OH B: acetonitrile + 5 mMNH₄OH Flow Rate 1.0 mL/min Gradient 2 to 98% B in 1.4 min

UPLC-MS 6:

Column Ascentis ® Express C18 2.7 μm Column Dimension 2.1 × 50 mm ColumnTemperature 80° C. Eluents A: water + 4.76% isopropanol + 0.05% FA +3.75 mM AA B: isopropanol + 0.05% FA Flow Rate 1.0 mL/min Gradient 1 to50% B in 1.4 min; 50-98% B in 0.3 min

UPLC-MS 7:

Column Acquity UPLC ® HSS T3 1.8 μm Column Dimension 2.1 × 50 mm ColumnTemperature 60° C. Eluents A: water + 0.05% formic acid + 3.75 mMammonium acetate B: acetonitrile + 0.04% FA Flow Rate 1.0 mL/minGradient 2 to 98% B in 1.4 min

UPLC-MS 8:

Column Acquity UPLC ® HSS T3 1.8 μm Column Dimension 2.1 × 50 mm ColumnTemperature 60° C. Eluents A: water + 0.05% formic acid + 3.75 mMammonium acetate B: acetonitrile + 0.04% FA Flow Rate 1.0 mL/minGradient 5 to 98% B in 1.4 min

UPLC-MS 9:

Column XBridge ® BEH ™ C18 2.5 μm Column Dimension 2.1 × 50 mm ColumnTemperature 80° C. Eluents A: water + 5 mM NH₄OH B: acetonitrile + 5 mMNH₄OH Flow Rate 1.0 mL/min Gradient 2 to 98% B in 9.4 min

UPLC-MS 10:

Column CORTECS ™ C18+ 2.7 μm, Column Dimension 2.1 × 50 mm ColumnTemperature 80° C. Eluents A: water + 0.05% FA + 3.75 mM AA B:isopropanol + 0.05% FA Flow Rate 1.0 mL/min Gradient concave from 1 to98% B in 1.4 min

UPLC-MS 11:

Instrument Shimadzu NEXERA UPLC PDA with Shimadzu LCMS 2020 as MSDColumn Mercury MS Synergi C12 2.5 μm Column Dimension 20 × 4.0 mm ColumnTemperature 40° C. Eluents A: water + 0.1% FA B: acetonitrile Flow Rate2.0 mL/min Gradient Time/% B: 0.01/5, 0.5/5, 1.0/95, 1.5/95, 2.0/5,3.0/5

UPLC-MS 12:

Instrument Agilent 1200 HPLC PDA with AB Sciex API2000 TQ as MSD ColumnMercury MS Synergi C12 2.5 μm Column Dimension 20 × 4.0 mm ColumnTemperature 30° C. Eluents A: water + 0.1% FA B: acetonitrile Flow Rate2.0 mL/min Gradient Time/% B: 0.01/30, 0.5/30, 1.0/95, 2.4/95, 2.5/30,3.0/30

UPLC-MS 13:

Instrument Agilent 1200 HPLC PDA with AB Sciex API3200 QTRAP as MSDColumn Kinetex EVO C18 2.6 μm Column Dimension 50 × 4.6 mm ColumnTemperature 30° C. Eluents A: water + 0.1% FA B: acetonitrile + 0.1% FAFlow Rate 1.5 mL/min Gradient Time/B: 0/20, 0.2/50, 1/95, 2.7/95,2.8/20, 4/20

UPLC-MS 14:

Column Acquity UPLC ® HSS T3 1.8 μm Column Dimension 2.1 × 100 mm ColumnTemperature 60° C. Eluents A: water + 0.05% formic acid + 3.75 mMammonium acetate B: acetonitrile + 0.04% FA Flow Rate 1.0 mL/minGradient 5 to 98% B in 9.4 min

HPLC Methods:

HPLC 1:

Instrument Agilent 1100 series with PDA Detector Column Kinetex C-18, 5μm Column Dimension 150 × 4.6 mm Column Temperature 40° C. Eluents A:water + 0.01% TFA B: acetonitrile Flow Rate 1.0 mL/min Gradient Time/B:0/30, 2/40, 5/90, 8/100, 10/100, 11/30, 12/30

HPLC 2:

Instrument Acquity Arc Waters UHPLC With PDA Dectector(2998 PDA) ColumnKinetex EO, 2.6 μm Column Dimension 100 × 4.6 mm Column Temperature 40°C. Eluents A: water + 0.01% TFA B: acetonitrile Flow Rate 1.0 mL/minGradient Time/B: 0/5, 2/5, 6/70, 10/100, 13/100, 13.5/5, 15/5

HPLC 3:

Instrument Agilent 1260 HPLC Column Agilent Poroshell 120 C18, 2.7 μmColumn Dimension 4.6 × 50 mm Column Temperature 40° C. Eluents A:water + 0.01% TFA B: acetonitrile + 0.01% TFA Flow Rate 1.2 mL/minGradient 0% B to 50% B in 5 min, hold 2 min

HPLC 4:

Instrument Agilent 1260 Column Agilent Poroshell 120 EC-C18, 2.7 μmColumn Dimension 4.6 × 50 mm Column Temperature 40° C. Eluents A:water + 0.1% TFA B: acetonitrile + 0.1% TFA Flow Rate 1.2 mL/minGradient 5% B to 95% B in 5 min, hold 2 min

HPLC 5:

Instrument Agilent 1260 HPLC Column InertSustain C18, 5 μm ColumnDimension 4.6 × 150 mm Column Temperature 30° C. Eluents A: water + 5mmol (NH₄)₂CO₃ B: acetonitrile Flow Rate 1.0 mL/min Gradient 10% B to90% B in 8 min, hold 2 min

HPLC 6:

Instrument Agilent 1260 infinity series HPLC system with DAD/ELSD ColumnAtlantis dC18, 5 μm Column Dimension 4.6 × 250 mm Column Temperature 25°C. Eluents A: water + 0.1% TFA B: acetonitrile Flow Rate 1.0 mL/minGradient 10% B to 100% B in 15 min, hold 5 min

Chiral HPLC Methods:

C-HPLC 1:

Instrument: Agilent 1260 Infinity II series with PDA Detector Injection:3 μL Mobile phase: A: hexane B: 0.1% HCOOH in EtOH Flow rate: 20 mL/minColumn: CELLULOSE 4 (150 × 4.6 mm, 5 μm) Detection UV: 210 nm Gradient:Isocratic: 50:50

C-HPLC 2:

Instrument: Agilent 1260 Infinity II series with PDA Detector Injection:10 μL Mobile phase: A: hexane B: 0.1% HCOOH in EtOH Flow rate: 15 mL/minColumn: CELLULOSE 4 (150 × 4.6 mm, 5 μm) Detection UV: 210 nm Gradient:Isocratic: 50:50

C-HPLC 3:

Instrument: Agilent 1260 Infinity II series with PDA Detector Injectionvolume: 8 μL Mobile phase: A: hexane B: 0.1% HCOOH in EtOH Flow rate: 1mL/min Column: CELLULOSE 4 (150 × 4.6 mm, 5 μm) Detection UV: 210, or254 nm Gradient: Isocratic: 50:50

C-HPLC 4:

Instrument: Agilent 1260 Infinity II series with PDA Detector Injectionvolume: 3 μL Mobile phase: A: hexane B: 0.1% HCOOH in EtOH Flow rate: 1mL/min Column: CELLULOSE 4 (150 × 4.6 mm, 5 μm) Detection UV: 210, or254 nm Gradient: Isocratic: 50:50

C-HPLC 5:

Instrument: Agilent 1260 Infinity II series with PDA Detector Injectionvolume: 2 μL Mobile phase: A: hexane B: 0.1% TFA in EtOH Flow rate: 1mL/min Column: CELLULOSE 4 (150 × 4.6 mm, 5 μm) Detection UV: 210, or254 nm Gradient: Isocratic: 50:50

C-HPLC 6:

Instrument: Agilent 1260 Infinity II series with PDA Detector Injectionvolume: 3 μL Mobile phase: A: hexane B: 0.1% TFA in EtOH Flow rate: 1mL/min Column: CELLULOSE 4 (150 × 4.6 mm, 5 μm) Detection UV: 210, or254 nm Gradient: Isocratic: 50:50

C-HPLC 7:

Instrument: Waters UPC² MS Injection volume: 5 μL Mobile phase: A: 25%(MeOH + 0.1% NH₃); B: 75% scCO₂ isocractic Flow rate: 3 mL/min Column:Chiralpak IB-N 5 μm, 100 × 4.6 mm Detection UV: 190-400 nm OvenTemperature: 40° C.

C-HPLC 8:

Instrument: Waters UPC² MS Injection volume: 5 μL Mobile phase: A: 35%(MeOH + 0.1% NH₃); B: 65% scCO₂ isocratic Flow rate: 3 mL/min Column:Chiralpak IB-N 5 μm, 100 × 4.6 mm Detection: 190-400 nm OvenTemperature: 40° C.

C-HPLC 9:

Instrument: Waters UPC² Injection volume: 1 μL Mobile phase: A: CO₂; B:MeOH + 0.05% DEA Flow rate: 2.4 mL/min Column: ChiralPak IC, 3 μm, 100 ×4.6 mm Detection: 220 nm Column Temperature: 35° C. Back pressure: 100bar

C-HPLC 10:

Instrument: Waters UPC²-MS Injection volume: 5 μL Mobile phase:isocratic A = 35% (IPA + 0.1% NH₃) B = 65% scCO₂ Flow rate: 3 mL/minColumn: ChiralPak IB, 5 μm, 100 × 4.6 mm Detection: DAD 210-400 nm Backpressure: 1800 psi

C-HPLC 11:

Instrument: Waters Acquity UPC Injection volume: 5 μL Mobile phase: A:35% (MeOH + 0.05% NH₃); B: 65% scCO₂ Flow rate: 3 mL/min Column:Chiralpak IB-N 5 μm, 100 × 4.6 mm Detection UV: 240 nm Oven Temperature:40° C.

C-HPLC 12:

Instrument: Waters Acquity UPC Injection volume: 5 μL Mobile phase: A:35% (MeOH + 0.05% NH₃); B: 65% scCO₂ Flow rate: 3 mL/min Column:Chiralpak IB-N 5 μm, 100 × 4.6 mm Detection UV: 240 nm Oven Temperature:40° C.

C-HPLC 13:

Instrument: Waters UPC²-MS Injection volume: 5 μL Mobile phase:isocratic A = 25% (MeOH + 0.05% NH₃) B = 75% scCO₂ Flow rate: 3 mL/minColumn: ChiralPak IB, 5 μm, 100 × 4.6 mm Detection: DAD 210-400 nm Backpressure: 1800 psi

Preparative Methods:

Column Chromatography:

Column chromatography was run on silica gel using prepacked columns, asdetailed below, or using glass columns following standard flashchromatography methodology, unless otherwise stated.

System 1 Teledyne ISCO, CombiFlash Rf, CombiFlash Rf+ System 2 BiotageIsolera Column pre-packed RediSep Rf cartridges, or SNAP cartridgesSample onto Isolute, or on silica gel, or applied as solutionsadsorption

Supercritical Fluid Chromatography (SFC)

Purifications were achieved on a Waters Preparative SFC-100-MS systemwith ABSYS update, with a Waters 2998 Photodiode Array Detector and aWaters MS Single Quadrupole Detector.

SFC 1:

Instrument: WATERS SFC 100 with ABSYS update Mobile phase: A: CO₂, B:MeOH Flow rate: 150 mL/min MeOH + 30 mL/min CO₂, constant flow of 180mL/min Column: 250 × 30 Reprospher PEI 100A 5 um Temperature: 50° C.Back pressure: 100 bar Detection UV: 210-400 nm Gradient: 18% B to 26% Bin 6.86 min

SFC 2:

Instrument: WATERS SFC 100 with ABSYS update Mobile phase: A: CO₂, B:MeOH Flow rate: 150 mL/min MeOH + 30 mL/min CO₂, constant flow of 180mL/min Column: 100 × 30 Reprosil NH₂ 100A 3 μm Temperature: 50° C. Backpressure: 100 bar Detection UV: 210-400 nm Gradient: 34% B to 42% B in2.8 min

SFC 3:

Instrument: WATERS SFC 100 with ABSYS update Mobile phase: A: CO₂, B:MeOH Flow rate: 150 mL/min MeOH + 30 mL/min CO₂, constant flow of 180mL/min Column: 100 × 30 Reprosil NH₂ 100A 3μpm Temperature: 50° C. Backpressure: 100 bar Detection UV: 210-400 nm Gradient: 21% B to 29% B in2.8 min first run 25% B to 33% B in 2.8 min second run to improve thedetection

SFC 4:

Instrument: WATERS SFC 100 Mobile phase: A. CO2, B: MeOH Flow rate: 100mL/min Column: 250 × 30 Reprospher PEI 100A 5 μm Temperature: 32° C.Back pressure: 120 bar Detection UV: 210-400 nm Gradient: 40% B to 55% Bin 10.2 min

SFC 5:

Instrument: WATERS SFC 100 Mobile phase: A: CO₂, B: MeOH Flow rate: 100mL/min MeOH Column: 250 × 30 Reprospher PEI 100A 5 μm Temperature: 32°C. Back pressure: 120 bar Detection UV: 210-400 nm Gradient: 35% B to45% B in 10.3 min

SFC 6:

Instrument: WATERS SFC 100 with ABSYS update Mobile phase: A: CO₂, B:MeOH Flow rate: 150 mL/min MeOH + 30 mL/min CO₂, constant flow of 180mL/min Column: 100 × 30 Reprosil NH₂ 100A 3 μm Temperature: 50° C. Backpressure: 100 bar Detection UV: 210-400 nm Gradient: 31% B to 39% B in2.8 min

SFC 7:

Instrument: WATERS SFC 100 with ABSYS update Mobile phase: A: CO₂, B:MeOH Flow rate: 150 mL/min MeOH + 30 mL/min CO₂, constant flow of 180mL/min Column: 250 × 30 waters Torus 2-PIC 130A 5 μm Temperature: 50° C.Back pressure: 100 bar Detection UV: 210-400 nm Gradient: 26% B to 34% Bin 5.48 min

Reversed Phase HPLC:

RP-HPLC Basic 1:

System Gilson Column Waters X-Bridge Prep C18 OBD (100 mm × 30 mm), 5 μmEluents A: water + 7.3 mM NH₄OH, B: acetonitrile Flow rate 40 mL/min

RP-HPLC Basic 2:

System Agilent 1200 series, with single quad mass spectrometer ColumnXBRIDGE(150 mm × 20 mm), 5.0 μm Eluents A: 0.02% ammonia in water, B:acetonitrile Flow rate 20 mL/min

RP-HPLC Acidic 1:

System Gilson Column Waters SunFire Prep C18 OBD (100 mm × 30 mm), 5 μmEluents A: water + 0.1% TFA, B: acetonitrile Flow rate 40 mL/min

RP-HPLC Acidic 2:

System Agilent 1200 series, with single quad mass spectrometer ColumnLUNA C18 (250 mm × 19 mm), 4.0 μm Eluents A: 0.1% HCOOH in water, B:acetonitrile Flow rate 20 mL/min

RP-HPLC Acidic 3:

System Agilent 1200 series, with single quad mass spectrometer ColumnLUNA (250 mm × 21.2 mm), 5.0 μm Eluents A: 0.1% HCOOH in water, B:acetonitrile Flow rate 18 mL/ min

RP-HPLC Acidic 4:

System Agilent 1200 series, with single quad mass spectrometer ColumnLUNA (250 mm × 21.2 mm), 5.0 μm Eluents A: 0.1% HCOOH in water, B:acetonitrile Flow rate 20 mL/min

RP-HPLC Acidic 5:

System Agilent 1200 series, with single quad mass spectrometer ColumnLUNA Phenomenex (250 mm × 21.2 mm), 5.0 μm Eluents A: 0.1% HCOOH inwater, B: acetonitrile Flow rate 18 mL/min

RP-HPLC Acidic 6:

System Agilent 1200 series, with single quad mass spectrometer ColumnAtlantis (250 mm × 19 mm), 5.0 μm Eluents A: 0.1% HCOOH in water, B:acetonitrile:MeOH Flow rate 18 mL/min

RP-HPLC Acidic 7:

System Agilent 1200 series, with single quad mass spectrometer ColumnLUNA C18 (250 mm × 21.2 mm), 5.0 μm Eluents A: 0.1% HCOOH in water, B:acetonitrile Flow rate 20 mL/min

RP-HPLC Acidic 8:

System Agilent 1200 series, with single quad mass spectrometer ColumnAtlantis (250 mm × 19 mm), 5.0 μm Eluents A: 0.1% HCOOH in water, B:acetonitrile Flow rate 18 mL/min

RP-HPLC Acidic 9:

System Agilent 1200 series, with single quad mass spectrometer ColumnAtlantis (250 mm × 21.2 mm), 5.0 μm Eluents A: 0.1% HCOOH in water, B:acetonitrile Flow rate 20 mL/min

RP-HPLC Acidic 10:

System Agilent 1200 series, with single quad mass spectrometer ColumnAtlantis (250 mm × 21.2 mm), 5.0 μm Eluents A: 0.1% HCOOH in water, B:acetonitrile Flow rate 18 mL/min

RP-HPLC Acidic 11:

System Agilent 1200 series, with single quad mass spectrometer ColumnLUNA OMEGA (250 mm × 21.2 mm), 5.0 μm Eluents A: 0.1% HCOOH in water, B:acetonitrile:MeOH (1:1) Flow rate 20 mL/min

RP-HPLC Acidic 12:

System Agilent 1200 series, with single quad mass spectrometer ColumnLUNA C18 (250 mm × 21.2 mm), 5.0 μm Eluents A: 0.1% HCOOH in water, B:acetonitrile Flow rate 18 mL/min

RP-HPLC Acidic 13:

System Gilson Column Nucleodur C18 (21 mm × 250 mm) Eluents A: water +0.1% COOH, B: acetonitrile + 0.1% COOH Flow rate 40 mL/min

RP-HPLC Acidic 14:

System Teledyne/Isco AccqPrep HP150 prep Column 50 × 100 Xbridge C18 (50mm × 100 mm), 5 μm Eluents A: water + 0.1% TFA, B: acetonitrile Flowrate 100 mL/min

RP-HPLC Neutral 1:

System Agilent 1200 series, with single quad mass spectrometer ColumnKINETEX (150 mm × 21.2 mm), 5 μm Eluents A: water, B: acetonitrile Flowrate 20 mL/min

RP-HPLC Neutral 2:

System Agilent 1200 series, with single quad mass spectrometer ColumnATLANTIS (250 mm × 21.2 mm), 5 μm Eluents A: water, B: acetonitrile Flowrate 17 mL/min

RP-HPLC Neutral 3:

System Agilent 1200 series, with single quad mass spectrometer ColumnLUNA C18 (250 mm × 21.2 mm), 5 μm Eluents A: water, B: acetonitrile Flowrate 20 mL/min

RP-HPLC Neutral 4:

System Agilent 1200 series, with single quad mass spectrometer ColumnLUNA Phenomenex (250 mm × 21.2 mm), 5 μm Eluents A: water, B:acetonitrile Flow rate 18 mL/min

RP-HPLC Neutral 5:

System Agilent 1200 series, with single quad mass spectrometer ColumnLUNA (250 mm × 21.2 mm) Eluents A: water, B: acetonitrile Flow rate 20mL/min

Preparation of Compounds

The following examples are intended to illustrate the invention and arenot to be construed as being limitations thereon. Temperatures are givenin degrees Celsius. If not mentioned otherwise, all evaporations areperformed under reduced pressure, typically between about 15 mm Hg and100 mm Hg (=20-133 mbar). Abbreviations used are those conventional inthe art.

All starting materials, building blocks, reagents, acids, bases,dehydrating agents, solvents, and catalysts utilized to synthesize thecompounds of the present invention are either commercially available orcan be produced by organic synthesis methods known to one of ordinaryskill in the art. Further, the compounds of the present invention can beproduced by organic synthesis methods known to one of ordinary skill inthe art as shown in the following examples.

The structures of all final products, intermediates and startingmaterials are confirmed by standard analytical spectroscopiccharacteristics, e.g., MS, IR or NMR. The absolute stereochemistry ofcertain isomers has been determined by analyses of X-ray crystalstructures of complexes in which the respective compounds are bound toWRN or by small molecule X-ray crystal structures of a precursor of thefinal compound.

Amines synthetized via acidic deprotection of the Boc-precursor wereoften obtained as HCl or TFA salt. The corresponding free base can beisolated by partitioning between DCM and aq sat NaHCO3 as described forIntermediate Y.

General Conditions:

Mass spectra were acquired on LC-MS systems using electrospray, chemicaland electron impact ionization methods with a range of instruments ofthe following configurations: Waters Acquity UPLC with Waters SQdetector, Shimadzu NEXERA UPLC PDA with Shimadzu LCMS 2020 as MSD,Agilent 1200 HPLC PDA with AB Sciex API2000 TQ as MSD and Agilent 1200HPLC PDA with AB Sciex API3200 QTRAP as MSD. [M+H]⁺ refers to theprotonated molecular ion of the chemical species.

NMR spectra were run with Bruker Ultrashield™400 (400 MHz), BrukerUltrashield™400 Plus (400 MHz), Bruker Ultrashield™600 (600 MHz) andBruker Ascend™400 (400 MHz) spectrometers, all with and withouttetramethylsilane as an internal standard. Chemical shifts (d-values)are reported in ppm downfield from tetramethylsilane, spectra splittingpattern are designated as singlet (s), doublet (d), triplet (t),multiplet, unresolved or more overlapping signals (m), broad signal(br). Solvents are given in parentheses.

Celite: Celite^(R) (the Celite corporation)=filtering aid based ondiatomaceous earth

Phase separator: Biotage—Isolute phase separator—(Part number:120-1906-D for 15 mL,

Part number: 120-1908-F for 70 mL and Part number: 120-1909-J for 150mL)

SiliaMetS®Thiol: SiliCYCLE thiol metal scavenger—(Part number: R51030B,Loading: 1.31 mmol/g Particle Size: 40-63 μm)

ISOLUTE® Si-Thiol: Biotage thiol metal scavenger—(Part number:9180-0100, Loading: 1.3 mmol/g)

PL-BnSH MP-Resin: Agilent thiol metal scavenger—(Part number:PL3582-6689, 2.2 mmol/g 100A 150-1 kg)

ISOLUTE® Si-TMT: Biotage thiol metal scavenger—(Part number: 9538)

Smopex®-301: Alfa Aesar thiol metal scavenger (Part number: 45902)

PL-HCO₃ MP SPE cartridge (500 mg per 6 mL)—(Part number: PL3540-C603)

PL-HCO₃ MP SPE cartridge (100 mg per 6 mL)—(Part number: PL3540-A603)

Selected compounds were crystallized and further characterized. Theexperimental procedures are outlined in the examples, infra, and theinstrument and method descriptions are outlined below:

X-ray Powder Diffraction Instrument and Method, FIGS. 1 to 5 InstrumentModel Bruker D8 Discover new Detector VANTEC-500 Radiation (Wavelength)Cu (1.5418 Å) X-ray generator power 40 kV, 40 mA Step size, resolutionTime/step 60 s, steps 2 Scan range 9° to 36° (2Theta value) Scan time120 s Temperature Room temperature Source slit Optics-Goebel mirror-Cu,Microslit-10 height 1 mm, UBC-Collimator-G-10 width 1 mm

X-ray Powder Diffraction Instrument and Method, FIG. 7 Instrument ModelBruker D8 Advance Detector LynxEye (1D mode), open angle: 2.948°Radiation (Wavelength) CuKα (0.15418 nm) Monochromator Ni-filter X-raygenerator power 40 kV, 40 mA Step size 0.0164° (2theta) Time per step0.3 s Scan range 2°-40° 2theta Scan time 768 s Source slit Primary fixedilluminated sample size: 10 mm, secondary: open angle: 2.2°, axialsoller: 2.5°

Sodium Salt Formation:

The compound was suspended in tert-butanol. NaOH 0.1M (1 eq) was added.The mixture was stirred/sonicated at RT. If the suspension turned into aclear solution it was lyophilized. If the suspension was still turbid,water was added and the resulting solution was lyophilized. If no changehappened, NaOH 0.1M up to 2 eq in total was added until a clear solutionwas observed, which was then lyophilized. If the NMR of the resultingsolid still contained tert-butanol, the solid was dissolved in a smallamount of water and lyophilized again. The final sodium salts wereobtained as colourless powders. The amorphous state was confirmed byXRPD.

Synthetic Schemes

Typically, the compounds of formula (I) can be prepared according to theSchemes provided infra. The examples which outline specific syntheticroutes, and the generic schemes below provide guidance to the syntheticchemist of ordinary skill in the art, who will readily appreciate thatthe solvent, concentration, reagent, protecting group, order ofsynthetic steps, time, temperature, and the like can be modified asnecessary.

The schemes provided infra are intended to represent singlediastereomers/enantiomers as well as their isomeric mixtures. Separationof diastereomers/enantiomers may be performed according to techniquesdescribed herein.

The invention includes the novel processes described herein, and furtherincludes any variant of the present processes, in which an intermediateobtainable at any stage thereof is used as starting material and theremaining steps are carried out, or in which the starting materials areformed in situ under the reaction conditions, or in which the reactioncomponents are used in the form of their salts or optically purematerial. Compounds of the present invention and intermediates can alsobe converted into each other according to methods generally known tothose skilled in the art. In another aspect, the invention providesnovel intermediate compounds, as described herein.

Abbreviations

Abbreviation Description AA ammonium acetate ACN acetonitrile AcOHacetic acid AgBF₄ silver tetrafluoroborate ATCC American Type CultureCollection, Mannassas, Virginia aq aqueous BCl₃ boron trichloride BINAP2,2′-bis(diphenylphosphino)-1,1′-binaphthyl Boc tert-butyloxycarbonylBoc₂O di-tert-butyl decarbonate br broad signal brine saturated aqueoussodium chloride ° C. degrees Celsius CaCl₂ calcium chloride CO carbonmonoxide CO₂ carbon dioxide CPhos2-dicyclohexylphosphino-2′,6′-bis(N,N-dimethyl- amino)biphenyl CPhos PdG3 [(2-dicyclohexylphosphino-2′,6′-bis(N,N-dimethylamino)-1,1′-biphenyl)-2-(2′-amino- 1,1′-biphenyl)] palladium(II)methanesulfonate Cs₂CO₃ caesium carbonate DAST diethylaminosulfurtrifluoride DCM dichloromethane DIBAL-H diisobutylaluminium hydrideDIPEA N,N-diisopropylethylamine DMAP 4-dimethylaminopyridine DME1,2-dimethoxyethane DMF N,N-dimethylformamide DMSO dimethyl sulfoxideDMSO-d₆ hexadeuterodimethyl sulfoxide dppf1,1′-bis(diphenylphosphino)ferrocene EDC•HCl3-(ethyliminomethyleneamino)-N,N-dimethyl- propan-1-amine hydrochlorideEDCl 3-(ethyliminomethyleneamino)-N,N-dimethyl- propan-1-amine eeenantiomeric excess eq equivalent Et₃N triethylamine Et₂O diethyl etherEtOAc ethylacetate EtOH ethanol FA formic acid H₂O water HATU1-[Bis(dimethylamino)methylene]-1H-1,2,3- triazolo[4,5-b]pyridinium3-oxide hexafluorophosphate HBr hydrobromic acid HCl hydrochloric acidHCOOH formic acid Hg mercury HI hydroiodic acid HNO₃ nitric acid HOAt3H-[1,2,3]triazolo[4,5-b]pyridin-3-ol HOBt benzotriazol-1-ol HPLChigh-performance liquid chromatography H₃PO₄ phosphoric acid HV highvacuum Hz, MHz, GHz hertz, megahertz, gigahertz IPA isopropyl alcoholIPAc isopropyl acetate J coupling constant K₂CO₃ potassium carbonatekg/g/mg kilogram/gram/milligram KHMDS potassium bis(trimethylsilyl)amideKI potassium iodide KMnO₄ potassium permanganate KOH potassium hydroxideK₃PO₄ potassium phosphate L/mL/μL liter/milliliter/microliter LC-MSliquid chromatography and mass spectrometry LDA lithium diisopropylamideLiCl lithium chloride M/mM molar/millimolar mbar millibar MCC padmicrocrystalline cellulose MeOH methanol MeTHF2-methyloxolane/2-methyltetrahydrofuran MgSO₄ magnesium sulfate minminutes mm/nm/μm millimeter/nanometer/micrometer μM/mMmicromolar/millimolar μmol/mmol/mol micromol/millimol/mol MS massspectrometry MW, mw microwave m/z mass to charge ratio N normality NaClsodium chloride NaCN sodium cyanide NaH sodium hydride NaHCO₃ sodiumbicarbonate NaH₂PO₄ sodium dihydrogen phosphate NaHSO4 sodium bisulfateNaOH sodium hydroxide Na₂O₃S₂ sodium thiosulfate NaOtBu sodium2-methylpropan-2-olate Na₂SO₄ sodium sulfate NBS N-bromosuccinimide NH₃ammonia NH₄Cl ammonium chloride NH₄OH ammonium hydroxide (NH₄)₂CO₃ammonium carbonate NIS N-iodosuccinimide NMP 1-methylpyrrolidin-2-oneNMR nuclear magnetic resonance PdCl₂(PPh₃)₂bis(triphenylphosphine)palladium(II) dichloride Pd(dppf)Cl₂[1,1′-bis(diphenylphosphino)ferrocene]palladium(II) dichloridePd(dppf)Cl₂•DCM [1,1′-Bis(diphenylphosphino)ferrocene]palladium(II)dichloride, DCM adduct Pd(OAc)₂ 1palladium(II) acetate Pd(PPh₃)₄tetrakis(triphenylphosphine)palladium(0) PMa pressure maximum allowablePOCl₃ phosphoryl chloride PtO₂ platinum dioxide PyAOP(7-azabenzotriazol-1-yloxy)tripyrrolidinophospho- niumhexafluorophosphate QD once daily (quaque die) R_(f) retardation factorRM or rm reaction mixture RP reversed phase RPM revolutions per minuteRt retention time (if not indicated, in minutes) RT room temperatureRuPhos 2-dicyclohexylphosphino-2′,6′-diisopropoxybiphenyl s, d, dd, t, msingulet, doublet, doublet of doublets, triplet, multiplet sat saturatedscCO₂ supercritical CO₂ SEM β-(trimethylsilyl)ethoxymethyl SFCsupercritical fluid chromatography T₃P propanephosphonic acid anhydrideTBAF tetrabutylammonium fluoride TBME 2-methoxy-2-methylpropane TFAtrifluoroacetic acid THF tetrahydrofuran TLC thin-layer chromatographyTMSOTf trimethylsilyl trifluoromethanesulfonate UPLC ultra performanceliquid chromatography W watt XPhos Pd G2chloro(2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl)[2-(2′-amino-1,1′- biphenyl)]palladium(II) XPhos Pd G3(2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II) methanesulfonate XRPDX-Ray Powder Diffraction

Preparation of Final Compounds

Example 1:2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-5-ethyl-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate B) (630 mg, 1.11 mmol) was suspended in DMF (6 mL).4-chloro-3-hydroxypicolinic acid (384 mg, 2.21 mmol), DIPEA (967 μL,5.54 mmol), HOBt (299 mg, 2.21 mmol) and EDC.HCl (425 mg, 2.21 mmol)were added to the RM and it was stirred at RT for 12 hours. Water wasadded to the RM and the suspension was filtered. The resulting solid waspurified by reverse phase preparative HPLC (RP-HPLC acidic 5: 20 to 30%B in 2 min, 30 to 60% B in 8 min) to give the title compound.

LC-MS: Rt=1.09 min; MS m/z [M+H]⁺ 724.6/726.6, m/z [M−H]⁻ 722.3/724.3;UPLC-MS 1 LC-MS: Rt=5.33 min; MS m/z [M+H]⁺ 724.2/726.2, m/z [M−H]⁻722.3/724.2; UPLC-MS 2 ¹H NMR (400 MHz, DMSO-d₆) δ 10.83 (s, br, 1H),10.34 (s, 1H), 8.05 (m, 2H), 7.96 (d, J=2.1 Hz, 1H), 7.72 (dd, J=2.1 Hz,8.8 Hz, 1H), 7.55 (d, J=5.1 Hz, 1H), 5.21 (s, 2H), 4.53 (m, 1H), 3.66(m, 4H), 3.54 (m, 3H), 3.38 (m, 4H), 3.23 (m, 1H), 2.96 (m, 3H), 2.78(m, 1H), 2.60 (m, 1H), 1.16 (t, J=7.3 Hz, 3H)

Example 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate B) (200 mg, 352 μmol) was suspended in DMF (5 mL).Perfluorophenyl 3-hydroxypicolinate (Intermediate CT) (215 mg, 703 μmol)and Et₃N (97.0 μL, 703 μmol) were added and the RM was stirred at 70° C.for 3 hours. The RM was concentrated under reduced pressure. The crudeproduct was first purified by column chromatography (Silica gel column:Silica 12 g, eluent DCM:MeOH 100:0 to 90:10). Then a second purificationby reverse phase preparative HPLC (RP-HPLC acidic 9: 40 to 50% B in 2min, 50 to 55% B in 10 min) afforded the title compound.

LC-MS: Rt=0.98 min; MS m/z [M+H]⁺ 690.6/692.6, m/z [M−H]⁻ 688.4/690.3;UPLC-MS 1

LC-MS: Rt=4.84 min; MS m/z [M+H]⁺ 690.2/692.2 m/z [M−H]⁻ 688.3/690.3;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.37 (s, br, 1H), 10.34 (s, br, 1H), 8.05(m, 2H), 7.96 (d, J=2.1 Hz, 1H), 7.72 (dd, J=2.1 Hz, 8.7 Hz, 1H), 7.28(m, 2H), 5.21 (s, 2H), 4.53 (m, 1H), 3.66 (m, 4H), 3.46 (m, 3H), 3.38(m, 4H), 3.20 (m, 1H), 2.92 (m, 3H), 2.76 (m, 1H), 2.58 (m, 1H), 1.16(t, J=7.5 Hz, 3H)

Example 3:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(4-fluoro-3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-(4-(3-(benzyloxy)-4-fluoropicolinoyl)piperazin-1-yl)-5-ethyl-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

ToN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide.TFA(Intermediate B) (620 mg, 908 μmol) in DMF (3 mL) was added Et₃N (503μL, 3.63 mmol) followed by 3-(benzyloxy)-4-fluoropicolinic acid(Intermediate CU) (224 mg, 908 μmol), then HATU (380 mg, 999 μmol). TheRM was stirred at RT for 30 minutes. The RM was diluted with water (5mL) and the resulting suspension was stirred at RT for 90 minutes. Thesuspension was filtered. The cake was washed with water (20 mL) anddried under vacuum to give the title compound as an off-white solid.

LC-MS: Rt=1.21 min; MS m/z [M+H]⁺ 798.5/800.5, m/z [M−H]⁻ 796.5/798.5;UPLC-MS 1.

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(4-fluoro-3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

2-(6-(4-(3-(benzyloxy)-4-fluoropicolinoyl)piperazin-1-yl)-5-ethyl-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(617 mg, 773 μmol) was dissolved in HBr (48% aq) (1.00 mL, 8.84 mmol)and stood at RT for 3 hours. Then it stood in a stoppered flask in thefridge over the weekend. The RM was allowed to warm to RT then heated at35° C. with stirring for 140 minutes. The RM was neutralized to pH 6 byaddition of 1M aq NaOH and extracted with DCM (2×30 mL). The combinedorganic layers were dried over Na₂SO₄, filtered and concentrated underreduced pressure. The crude product was purified in 2 portions byreverse phase preparative HPLC (RP-HPLC acidic 1: 30 to 50% B in 20 minwith a plateau at 50% for 1 min and RP-HPLC acidic 1: 20 to 47% B in 20min with a plateau at 20% for 1 min). The product containing fractionswere combined and partitioned between DCM (30 mL) and aq sat NaHCO₃ (5mL). The organic layer was separated by filtration through a phaseseparator and concentrated under reduced pressure. The residue wasrecrystallized from MeOH/water to give the title compound as colourlesscrystals.

LC-MS: Rt=0.98 min; MS m/z [M+H]⁺ 708.5/710.5, m/z [M−H]⁻ 706.4/708.4;UPLC-MS 1 LC-MS: Rt=4.89 min; MS m/z [M+H]⁺ 708.2/710.2, m/z [M−H]⁻706.2/708.2; UPLC-MS 2 ¹H NMR (400 MHz, DMSO-d₆) δ 10.72 (s, br, 1H),10.33 (s, 1H), 8.08 (dd, J=5.3 Hz, 6.9 Hz, 1H), 8.04 (d, J=8.7 Hz, 1H),7.96 (d, J=2.1 Hz, 1H), 7.72 (dd, J=2.2 Hz, 8.8 Hz, 1H), 7.35 (dd, J=5.3Hz, 11.9 Hz, 1H), 5.21 (s, 2H), 4.52 (m, 1H), 3.66 (m, 4H), 3.45 (m,3H), 3.38 (m, 4H), 3.21 (m, 1H), 2.94 (m, 3H), 2.77 (m, 1H), 2.59 (m,1H), 1.16 (t, J=7.4 Hz, 3H)

Example 4:N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate C) (400 mg, 686 μmol) was suspended in DMF (5 mL).3-hydroxypicolinic acid (191 mg, 1.37 mmol), DIPEA (599 μL, 3.43 mmol),HOBt (185 mg, 1.37 mmol) and EDC.HCl (263 mg, 1.37 mmol) were added tothe RM and stirred at RT for 12 hours. Water was added to the RM and itwas filtered. The solid was purified by reverse phase preparative HPLC(RP-HPLC neutral 5: 10 to 20% B in 2 min, 20 to 45% B in 10 min) to givethe title compound.

LC-MS: Rt=1.00 min; MS m/z [M+H]⁺ 704.1/706.2, m/z [M−H]⁻ 702.2/704.2;UPLC-MS 1

LC-MS: Rt=5.01 min; MS m/z [M+H]⁺ 704.2/706.1, m/z [M−H]⁻ 702.3/704.3;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.37 (s, br, 1H), 10.07 (s, 1H), 8.06 (m,1H), 7.93 (s, 1H), 7.75 (s, 1H), 7.28 (m, 2H), 5.16 (s, 2H), 4.53 (m,1H), 3.65 (m, 4H), 3.46 (m, 3H), 3.39 (m, 4H), 3.20 (m, 1H), 2.94 (m,3H), 2.75 (m, 1H), 2.58 (m, 1H), 2.34 (s, 3H), 1.16 (t, J=7.1 Hz, 3H)

Example 5:2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

2-(5-Ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide.HCl(Intermediate D) (230 mg, 381 μmol) and DIPEA (333 μL, 1.91 mmol) weredissolved in DCM (10 mL) and then 3-hydroxypicolinoyl chloride(Intermediate CV) (90.0 mg, 572 μmol) was added at 0° C. and stirred for1 hour. 3-hydroxypicolinoyl chloride (Intermediate CV) (90.0 mg, 572μmol) was added again at 0° C. and stirred for 1 hour. The RM wasdiluted with DCM and washed with water, aq NaHCO₃ solution (2×20 mL),again with water and brine, dried over Na₂SO₄, filtered andconcentrated. The crude product was purified by reverse phasepreparative HPLC (RP-HPLC acidic 7: 30 to 40% B in 2 min, 40 to 50% B in9 min) to give the title compound as a pale brown solid.

LC-MS: Rt=0.96 min; MS m/z [M+H]⁺ 688.2, m/z [M−H]⁻ 686.3; UPLC-MS 1

LC-MS: Rt=4.81 min; MS m/z [M+H]⁺ 688.2, m/z [M−H]⁻ 686.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.35 (s, boad, 1H), 10.06 (s, br, 1H), 8.05(m, 1H), 7.77 (d, J=13.0 Hz, 1H), 7.66 (d, J=8.2 Hz, 1H), 7.28 (m, 2H),5.18 (s, 2H), 4.53 (m, 1H), 3.66 (m, 4H), 3.46 (m, 3H), 3.38 (m, 4H),3.20 (m, 1H), 2.92 (m, 3H), 2.76 (m, 1H), 2.58 (m, 1H), 2.33 (s, 3H),1.15 (t, J=7.2 Hz, 3H)

Example 6:2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-5-ethyl-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

2-(5-Ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate D) (220 mg, 388 μmol) was suspended in DMF (5 mL).4-chloro-3-hydroxypicolinic acid (101 mg, 582 μmol), DIPEA (339 μL, 1.94mmol), HOBt (105 mg, 777 μmol) and EDC.HCl (149 mg, 777 μmol) were addedto the RM and it was stirred at RT for 12 hours. Water was added to theRM and the precipitate was filtered off. The obtained solid was purifiedby reverse phase preparative HPLC (RP-HPLC acidic 5: 20 to 30% B in 2min, 30 to 60% B in 8 min) to give the title compound.

LC-MS: Rt=1.07 min; MS m/z [M+H]⁺ 722.3/724.3, m/z [M−H]⁻ 720.3/722.3;UPLC-MS 1

LC-MS: Rt=5.32 min; MS m/z [M+H]⁺ 722.2/724.2 m/z [M−H]⁻ 720.2/722.2;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.82 (s, br, 1H), 10.05 (s, 1H), 8.06 (d,J=5.1 Hz, 1H), 7.77 (d, J=12.8 Hz, 1H), 7.67 (d, J=8.1 Hz, 1H), 7.55 (d,J=4.8 Hz, 1H), 5.18 (s, 2H), 4.53 (m, 1H), 3.65 (m, 4H), 3.53 (m, 3H),3.38 (m, 4H), 3.24 (m, 1H), 2.93 (m, 3H), 2.78 (m, 1H), 2.60 (m, 1H),2.33 (s, 3H), 1.16 (t, J=7.1 Hz, 3H)

Example 7:N-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

N-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide.HCl(Intermediate E) (300 mg, 481 μmol) and DIPEA (420 μL, 2.41 mmol) weredissolved in DCM (15 mL) and then 3-hydroxypicolinoyl chloride(Intermediate CV) (152 mg, 962 μmol) was added at 0° C. and stirred for2 hours. 3-hydroxypicolinoyl chloride (Intermediate CV) (152 mg, 962μmol) was added again and continued stirring for 14 hours. The RM wasdiluted with DCM and washed with water, aq NaHCO₃ solution (2×20 mL),again with water and brine, dried over Na₂SO₄, filtered andconcentrated. The crude product was purified by column chromatography(Silica gel column: Silica 12 g, eluent DCM:MeOH 100:0 to 99:1). Theproduct containing fractions were concentrated, then further purified byreverse phase preparative HPLC (RP-HPLC acidic 7: 40 to 50% B in 2 min,50 to 60% B in 8 min) to give the title compound as an off white solid.

LC-MS: Rt=1.01 min; MS m/z [M+H]⁺ 708.4/710.4, m/z [M−H]⁻ 706.4/708.4;UPLC-MS 1

LC-MS: Rt=5.11 min; MS m/z [M+H]⁺ 708.2/710.1 m/z [M−H]⁻ 706.2/708.2;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.44 (s, 1H), 10.37 (s, 1H), 8.07 (m, 2H),8.01 (d, J=7.3 Hz, 1H), 7.28 (m, 2H), 5.25 (s, 2H), 4.53 (m, 1H), 3.65(m, 4H), 3.46 (m, 3H), 3.37 (m, 4H), 3.22 (m, 1H), 2.93 (m, 3H), 2.76(m, 1H), 2.58 (m, 1H), 1.15 (t, J=7.4 Hz, 3H)

Example 8:2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To the stirred solution of 3-hydroxypicolinic acid (143 mg, 1.03 mmol)in DMF (2.5 mL) were added EDC.HCl (197 mg, 1.03 mmol) and HOBt (13.0mg, 1.03 mmol) at RT. In another flask2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide.HCl(Intermediate F) (300 mg, 513 μmol) in DMF (2.5 mL) was mixed with DIPEA(537 μL, 3.08 mmol) at RT. This solution was added to the first RM at RTand stirred for 12 hours. The RM was concentrated, water was added andit was extracted with EtOAc. The organic layer was washed with aq NaHCO₃solution (2 times), dried over Na₂SO₄ and concentrated under reducedpressure. The crude product was purified by reverse phase preparativeHPLC (RP-HPLC neutral 4: 30 to 40% B in 2 min, 40 to 50% B in 8 min) togive the title compound.

LC-MS: Rt=0.92 min; MS m/z [M+H]⁺ 670.4, m/z [M−H]⁻ 668.3; UPLC-MS 1

LC-MS: Rt=4.64 min; MS m/z [M+H]⁺ 670.3, m/z [M−H]⁻ 668.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.40 (s, 1H, br), 9.98 (s, 1H), 8.05 (m,1H), 7.70 (d, J=8.4 Hz, 1H), 7.62 (s, br, 1H), 7.53 (d, J=8.3 Hz, 1H),7.28 (m, 2H), 5.14 (s, 2H), 4.53 (m, 1H), 3.66 (m, 4H), 3.46 (m, 3H),3.39 (m, 4H), 3.20 (m, 1H), 2.95 (m, 3H), 2.76 (m, 1H), 2.58 (m, 1H),2.33 (s, 3H), 1.16 (t, J=7.1 Hz, 3H)

Example 9:2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-5-ethyl-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

4-Chloro-3-hydroxypicolinic acid (120 mg, 692 μmol) was dissolved in DMF(3 mL) and then2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide.HCl(Intermediate F) (270 mg, 462 μmol), EDC.HCl (177 mg, 923 μmol), DIPEA(403 μL, 2.31 mmol) and HOBt (125 mg, 923 μmol) were added at 0° C. andstirred at RT for 14 hours. The RM was diluted with water and extractedwith 5% MeOH in DCM and washed with aq sat NaHCO₃ and brine. Thecombined organic layers were dried over Na₂SO₄ and concentrated underreduced pressure. The crude product was purified by reverse phasepreparative HPLC (RP-HPLC acidic 7: 30 to 40% B in 2 min, 40 to 60% B in8 min) to give the title compound.

LC-MS: Rt=1.02 min; MS m/z [M+H]⁺ 704.2/706.2, m/z [M−H]⁻ 702.2/704.2;UPLC-MS 1

LC-MS: Rt=5.05 min; MS m/z [M+H]⁺ 704.2/706.2, m/z [M−H]⁻ 702.3/704.2;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.85 (s, br, 1H), 9.99 (s, 1H), 8.03 (d,J=5.1 Hz, 1H), 7.70 (d, J=8.4 Hz, 1H), 7.62 (s, br, 1H), 7.53 (m, 2H),5.14 (s, 2H), 4.53 (m, 1H), 3.66 (m, 4H), 3.53 (m, 3H), 3.39 (m, 4H),3.24 (m, 1H), 2.95 (m, 3H), 2.77 (m, 1H), 2.60 (m, 1H), 2.34 (s, 3H),1.17 (t, J=7.1 Hz, 3H)

Example 10:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate G) (900 mg, 1.58 mmol) was suspended in DMF (5 mL).Perfluorophenyl 3-hydroxypicolinate (Intermediate CT) (964 mg, 3.16mmol) and Et₃N (438 μL, 3.16 mmol) were added and the RM was stirred at70° C. for 3 hours. The RM was concentrated under reduced pressure. Thecrude product was purified by column chromatography (Silica gel column:Silica 24 g, eluent DCM:MeOH 100:0 to 99:1) and recrystallized usingisopropanol to give the title compound.

LC-MS: Rt=0.85 min; MS m/z [M+H]⁺ 691.4/693.4, m/z [M−H]⁻ 689.5/691.5;UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.52 (s, 1H), 10.36 (s, 1H), 8.54 (d, J=8.3Hz, 1H), 8.06 (m, 1H), 7.96 (d, J=8.5 Hz, 1H), 7.28 (m, 2H), 5.25 (s,2H), 4.53 (m, 1H), 3.65 (m, 4H), 3.46 (m, 3H), 3.37 (m, 4H), 3.20 (m,1H), 2.94 (m, 3H), 2.76 (m, 1H), 2.58 (m, 1H), 1.15 (t, J=7.1 Hz, 3H)

Example 11:2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide

To the stirred solution of 3-hydroxypicolinic acid (101 mg, 724 μmol),EDC.HCl (139 mg, 724 μmol) and HOBt (98.0 mg, 724 μmol) in DMF (3 mL)was added2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide(Intermediate H) (200 mg, 362 μmol) and then DIPEA (379 μL, 2.17 mmol)at RT. This RM was stirred at RT for 16 hours. The reaction wasconcentrated under reduced pressure and water was added. The resultantbrown solid was filtered and dried under vacuum. The crude product waspurified by reverse phase preparative HPLC (RP-HPLC neutral 1: 25 to 35%B in 2 min, 35 to 50% B in 9 min) to give the title compound.

LC-MS: Rt=0.95 min; MS m/z [M+H]⁺ 674.6, m/z [M−H]⁻ 672.4; UPLC-MS 1

LC-MS: Rt=4.68 min; MS m/z [M+H]⁺ 674.2, m/z [M−H]⁻ 672.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.58 (s, br, 2H), 8.21 (m, 1H), 8.04 (m,1H), 7.79 (d, J=10.9 Hz, 1H), 7.57 (d, J=8.7 Hz, 1H), 7.27 (m, 2H), 5.19(s, 2H), 4.53 (m, 1H), 3.65 (m, 4H), 3.46 (m, 3H), 3.37 (m, 4H), 3.20(m, 1H), 2.92 (m, 3H), 2.76 (m, 1H), 2.57 (m, 1H), 1.14 (t, J=7.2 Hz,3H)

Example 12:N-(4-chloro-2-methyl-5-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

3-Hydroxypicolinic acid (84.0 mg, 605 μmol) was dissolved in DMF (8 mL)and thenN-(4-chloro-2-methyl-5-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide.HCl(Intermediate 1) (250 mg, 404 μmol), EDC.HCl (116 mg, 605 μmol), DIPEA(423 μL, 2.42 mmol) and HOBt (82.0 mg, 605 μmol) were added at 0° C. andstirred at RT for 16 hours. The RM was partially concentrated anddiluted with water, extracted with EtOAc, washed with aq sat NaHCO₃ andbrine and the combined organic layers were dried over Na₂SO₄, filteredand concentrated under reduced pressure. The crude product was sonicatedin ACN and MeOH (1:1) (5 mL) and then filtered. The obtained solid waswashed with EtOAc and pentane and dried to give the title compound as anoff-white solid.

LC-MS: Rt=1.00 min; MS m/z [M+H]⁺ 704.6/706.5, m/z [M−H]⁻ 702.4/704.4;UPLC-MS 1

LC-MS: Rt=4.94 min; MS m/z [M+H]⁺ 704.2/706.2, m/z [M−H]⁻ 702.3/704.3;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.41 (s, br, 1H), 10.07 (s, br, 1H), 8.04(m, 1H), 7.94 (s, 1H), 7.66 (s, 1H), 7.27 (m, 2H), 5.12 (s, 2H), 4.53(m, 1H), 3.65 (m, 4H), 3.46 (m, 3H), 3.39 (m, 4H), 3.20 (m, 1H), 2.94(m, 3H), 2.75 (m, 1H), 2.58 (m, 1H), 2.33 (s, 3H), 1.16 (t, J=7.4 Hz,3H)

Example 13:rac-N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Rac-N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide.HCl(Intermediate K) (360 mg, 566 μmol), EDC.HCl (163 mg, 850 μmol),3-hydroxypicolinic acid (118 mg, 850 μmol) and HOBt (115 mg, 850 μmol)were dissolved in DMF (5 mL) and then DIPEA (594 μL, 3.40 mmol) wasadded at 0° C. and stirred at RT for 16 hours. The RM was diluted withwater and extracted with EtOAc and washed with aq sat NaHCO₃ and brineand the combined organic layers were dried over Na₂SO₄, filtered andconcentrated under reduced pressure. The crude product was purifiedtwice by column chromatography (2× Silica gel column: Silica 12 g,eluent DCM:MeOH 100:0 to 95:5) to give the title compound as an offwhite solid.

LC-MS: Rt=1.08 min; MS m/z [M+H]⁺ 720.1/722.0 m/z [M−H]⁻ 718.3/720.2;UPLC-MS 1

LC-MS: Rt=5.42 min; MS m/z [M+H]⁺ 720.1/722.1, m/z [M−H]⁻ 718.3/720.3;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.38 (s, 1H), 10.08 (s, 1H), 8.06 (m, 1H),7.92 (s, 1H), 7.76 (s, 1H), 7.29 (m, 2H), 5.15 (s, 2H), 4.75 (d, br, J,47.5 Hz, 1H), 4.53 (m, 1H), 3.73 (m, 1H), 3.58 (m, 2H), 3.44 (m, 3H),3.23 (m, 1H), 2.93 (m, 3H), 2.76 (m, 1H), 2.58 (m, 1H), 2.35 (s, 3H),1.81 (m, 4H), 1.52 (m, 1H), 1.16 (t, J=7.2 Hz, 3H)

Example 14:rac-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Rac-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide.HCl(Intermediate L) (300 mg, 499 μmol) was dissolved in DMF (4 mL).3-hydroxypicolinic acid (174 mg, 1.25 mmol), EDC.HCl (239 mg, 1.25mmol), HOBt (169 mg, 1.25 mmol) and DIPEA (436 μL, 2.50 mmol) were addedat 0° C. and the RM was stirred at RT for 24 hours. The RM was dilutedwith water and extracted with 5% MeOH in DCM, washed with aq sat NaHCO₃,brine and the organic layer was dried and concentrated under reducedpressure. The crude product was purified by reverse phase preparativeHPLC (RP-HPLC acidic 6: 40 to 50% B in 2 min, 50 to 70% B in 8 min) togive the title compound.

LC-MS: Rt=1.01 min; MS m/z [M+H]⁺ 686.2, m/z [M−H]⁻ 684.3; UPLC-MS 1

LC-MS: Rt=5.05 min; MS m/z [M+H]⁺ 686.2, m/z [M−H]⁻ 684.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.32 (s, br, 1H), 9.99 (s, 1H), 8.06 (m,1H), 7.69 (d, J=8.8 Hz, 1H), 7.62 (s, br, 1H), 7.53 (d, J=8.2 Hz, 1H),7.28 (m, 2H), 5.13 (s, 2H), 4.75 (d, br, J=48.2 Hz, 1H), 4.53 (m, 1H),3.72 (m, 1H), 3.58 (m, 2H), 3.43 (m, 3H), 3.21 (m, 2H), 2.93 (m, 3H),2.75 (m, 1H), 2.58 (m, 1H), 2.34 (s, 3H), 1.82 (m, 3H), 1.53 (m, 1H),1.16 (t, J=7.4 Hz, 3H)

Example 14a:(R)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamideand Example 14b:(S)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Chiral separation ofrac-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide:

Preparative chiral HPLC (instrument: Agilent 1200 series, with singlequad mass spectrometer; column: LUX CELLULOSE-4, 250 mm×21.2 mm; eluent:A=hexane, B=0.1% HCOOH in MeOH:EtOH 1:1; flow rate: 15.0 mL/min;detection: 210 nm; injection volume: 0.9 mL; Gradient: isocratic70(A):30(B)).

Example 14a: The first eluting stereoisomer was stirred in Et₂O (20 mL),filtered and the resultant solid was dried under vacuum to give thetitle compound.

Chiral HPLC (C-HPLC 3): Rt=6.17 min

LC-MS: Rt=1.01 min; MS m/z [M+H]⁺ 686.2, m/z [M−H]⁻ 684.3; UPLC-MS 1

LC-MS: Rt=5.05 min; MS m/z [M+H]⁺ 686.2, m/z [M−H]⁻ 684.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.36 (s, br, 1H), 10.00 (s, 1H), 8.05 (m,1H), 7.69 (d, J=8.5 Hz, 1H), 7.62 (s, br, 1H), 7.53 (d, J=8.4 Hz, 1H),7.28 (m, 2H), 5.13 (s, 2H), 4.85-4.65 (d, br, J=47.5 Hz, 1H), 4.53 (m,1H), 3.72 (m, 1H), 3.65-3.35 (m, 6H), 3.20 (m, 1H), 2.94 (m, 3H), 2.75(m, 1H), 2.57 (m, 1H), 2.34 (s, 3H), 1.81 (m, 3H), 1.52 (m, 1H), 1.16(t, J=7.3 Hz, 3H)

Example 14b: The second eluting stereoisomer was purified by reversephase preparative HPLC (RP-HPLC acidic 5: 20 to 30% B in 2 min, 30 to60% B in 8 min) to give the title compound.

Chiral HPLC (C-HPLC 4): Rt=8.20 min

LC-MS: Rt=1.02 min; MS m/z [M+H]⁺ 686.4, m/z [M−H]⁻ 684.4; UPLC-MS 1

LC-MS: Rt=5.12 min; MS m/z [M+H]⁺ 686.3, m/z [M−H]⁻ 684.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.43 (s, br, 1H), 10.00 (s, 1H), 8.05 (m,1H), 7.69 (d, J=8.5 Hz, 1H), 7.62 (s, br, 1H), 7.53 (d, J=8.5 Hz, 1H),7.28 (m, 2H), 5.13 (s, 2H), 4.85-4.65 (d, br, J=47.7 Hz, 1H), 4.53 (m,1H), 3.74 (m, 1H), 3.65-3.35 (m, 6H), 3.21 (m, 1H), 2.94 (m, 3H), 2.75(m, 1H), 2.58 (m, 1H), 2.34 (s, 3H), 1.78 (m, 3H), 1.54 (m, 1H), 1.16(t, J=7.4 Hz, 3H)

Example 15:rac-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Rac-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate M) (160 mg, 274 μmol) was suspended in DMF (2 mL) and3-hydroxypicolinic acid (95.0 mg, 684 μmol), DIPEA (239 μL, 1.37 mmol),HOBt (92.0 mg, 684 μmol) and EDC.HCl (131 mg, 684 μmol) were added tothe RM and it was stirred at RT for 12 hours. Water was added to the RM,the precipitate was filtered off and the obtained crude product waspurified by reverse phase preparative HPLC (RP-HPLC neutral 2: 30 to 40%B in 2 min, 40 to 75% B in 9 min) to give the title compound.

LC-MS: Rt=1.06 min; MS m/z [M+H]⁺ 706.3/708.2, m/z [M−H]⁻ 704.3/706.4;UPLC-MS 1

LC-MS: Rt=5.34 min; MS m/z [M+H]⁺ 706.2/708.1, m/z [M−H]⁻ 704.3/706.3;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.36 (2s, 2H), 8.04 (m, 2H), 7.96 (s, br,1H), 7.72 (d, J=8.7 Hz, 1H), 7.28 (m, 2H), 5.20 (s, 2H), 4.85-4.5 (d,br, J=48.6 Hz, 1H), 4.53 (m, 1H), 3.72 (m, 1H), 3.65-3.35 (m, 5H), 3.23(m, 2H), 2.94 (m, 3H), 2.75 (m, 1H), 2.57 (m, 1H), 1.77 (m, 3H), 1.52(m, 1H), 1.15 (t, J=7.2 Hz, 3H)

Example 15a:((R)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide)or((S)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide)and Example 15b:((R)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide)or((S)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide)

Chiral separation ofrac-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide:

Preparative chiral HPLC (instrument: SEPIATEC SFC100; column: OVEN3Chiralpak IB-N 250×30 mm 5 μm; eluent: A: 28% [MeOH+0.1% NH₃] B: 72%scCO₂; flow rate: 90.0 mL/min; detection: 236 nm; injection volume: 0.30mL; Gradient: isocratic A: 28%, B: 72% scCO₂

Example 15a: The first eluting stereoisomer was concentrated underreduced pressure at 35° C. to give a white solid.

Chiral HPLC (C-HPLC 7): Rt=3.33 min, 99% ee

LC-MS: Rt=1.05 min; MS m/z [M+H]⁺ 706.3/708.3, m/z [M−H]⁻ 704.3/706.3;UPLC-MS 3

¹H NMR (600 MHz, DMSO-d₆) δ 10.35 (2s, 2H), 8.05 (m, 1H), 8.03 (d, J=8.8Hz, 1H), 7.96 (d, J=2.1 Hz, 1H), 7.71 (dd, J=2.2 Hz, 8.7 Hz, 1H), 7.28(m, 2H), 5.20 (s, 2H), 4.80-4.65 (d, br, J=46.9 Hz, 1H), 4.53 (m, 1H),3.72 (m, 1H), 3.65-3.35 (m, 5H), 3.29 (m, 1H), 3.19 (m, 1H), 2.93 (m,3H), 2.75 (m, 1H), 2.58 (m, 1H), 1.95-1.7 (m, 3H), 1.52 (m, 1H), 1.15(t, J=7.3 Hz, 3H)

Example 15b: The second eluting stereoisomer was concentrated underreduced pressure at 35° C. to give a beige solid.

Chiral HPLC (C-HPLC 7): Rt=3.89 min, 94% ee

LC-MS: Rt=1.05 min; MS m/z [M+H]⁺ 706.3/708.3, m/z [M−H]⁻ 704.3/706.2;UPLC-MS 3

¹H NMR (600 MHz, DMSO-d₆) δ 10.35 (2s, 2H), 8.05 (m, 2H), 7.96 (d, J=2.1Hz, 1H), 7.71 (dd, J=2.1 Hz, 8.8 Hz, 1H), 7.28 (m, 2H), 5.20 (s, 2H),4.80-4.70 (d, br, J=47.8 Hz, 1H), 4.53 (m, 1H), 3.73 (m, 1H), 3.65-3.35(m, 5H), 3.28 (m, 1H), 3.20 (m, 1H), 2.93 (m, 3H), 2.75 (m, 1H), 2.58(m, 1H), 1.95-1.7 (m, 3H), 1.52 (m, 1H), 1.15 (t, J=7.3 Hz, 3H)

Example 16:rac-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Rac-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide.HCl(Intermediate N) (100 mg, 161 μmol) and DIPEA (140 μL, 803 μmol) weredissolved in DCM (5 mL) and then 3-hydroxypicolinoyl chloride(Intermediate CV) (50.6 mg, 321 μmol) was added at 0° C. and stirred for2 hours. The RM was diluted with DCM and washed with water and aq satNaHCO₃ (2×20 mL), washed with water and brine. The organic layer wasdried over Na₂SO₄, filtered and concentrated under reduced pressure. Thecrude product was purified by reverse phase preparative HPLC (RP-HPLCacidic 7: 30 to 40% B in 2 min, 40 to 50% B in 8 min) to give the titlecompound as a pale brown solid.

LC-MS: Rt=0.94 min; MS m/z [M+H]⁺ 707.6/709.6, m/z [M−H]⁻ 705.4/707.4;UPLC-MS 1

LC-MS: Rt=4.59 min; MS m/z [M+H]⁺ 707.2/709.2, m/z [M−H]⁻ 705.3/707.2;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.54 (s, 1H), 10.37 (s, 1H), 8.54 (d, J=8.3Hz, 1H), 8.06 (m, 1H), 7.96 (d, J=8.0 Hz, 1H), 7.28 (m, 2H), 5.25 (s,2H), 4.85-4.65 (d, br, J=47.2 Hz, 1H), 4.53 (m, 1H), 3.73 (m, 1H), 3.60(m, 1H), 3.46 (m, 4H), 3.23 (m, 2H), 2.94 (m, 3H), 2.75 (m, 1H), 2.57(m, 1H), 1.79 (m, 3H), 1.52 (m, 1H), 1.15 (t, J=7.3 Hz, 3H)

Example 16a:(R)—N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideand Example 16b:(S)—N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Chiral separation ofrac-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide:

Preparative chiral HPLC (instrument: Agilent 1200 series, with singlequad mass spectrometer; column: CELLULOSE-4, 250 mm×21.2 mm; eluent:A=hexane, B=0.1% HCOOH in MeOH:EtOH 1:1; flow rate: 18.0 mL/min;detection: 210 nm; injection volume: 0.9 mL; Gradient: isocratic70(A):30(B)). The chiral isomers which were separated and concentratedwere taken, washed with n-hexane, decanted, dried and analysed.

Example 16a: First eluting stereoisomer, off white solid.

Chiral HPLC (C-HPLC 5): Rt=6.189 min

LC-MS: Rt=0.93 min; MS m/z [M+H]⁺ 707.1/709.1, m/z [M−H]⁻ 705.3/705.2;UPLC-MS 1

LC-MS: Rt=4.60 min; MS m/z [M+H]⁺ 707.1/709.0, m/z [M−H]⁻ 705.3/705.2;UPLC-MS 2 ¹H NMR (400 MHz, DMSO-d₆) δ 10.63, (s, br, 2H), 8.53 (d, J=7.6Hz, 1H), 8.05 (m, 1H), 7.89 (d, J=7.6 Hz, 1H), 7.28 (m, 2H), 5.19 (s,2H), 4.85-4.65 (d, br, J=48.0 Hz, 1H), 4.53 (m, 1H), 3.71 (m, 1H),3.65-3.15 (m, 7H), 2.93 (m, 3H), 2.75 (m, 1H), 2.58 (m, 1H), 1.81 (m,3H), 1.52 (m, 1H), 1.15 (t, J=7.3 Hz, 3H)

Example 16b: Second eluting stereoisomer, off-white solid.

Chiral HPLC (C-HPLC 6): Rt=7.575 min

LC-MS: Rt=0.93 min; MS m/z [M+H]⁺ 707.1/709.0, m/z [M−H]⁻ 705.3/705.2;UPLC-MS 1

LC-MS: Rt=4.60 min; MS m/z [M+H]⁺ 707.1/709.0, m/z [M−H]⁻ 705.3/705.2;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.43, (s, broad, 2H), 8.54 (d, J=8.2 Hz,1H), 8.05 (m, 1H), 7.94 (d, J=8.2 Hz, 1H), 7.28 (m, 2H), 5.23 (s, 2H),4.85-4.65 (d, br, J=48.3 Hz, 1H), 4.53 (m, 1H), 3.71 (m, 1H), 3.60 (m,1H), 3.46 (m, 4H), 3.21 (m, 2H), 2.93 (m, 3H), 2.75 (m, 1H), 2.58 (m,1H), 1.81 (m, 3H), 1.52 (m, 1H), 1.15 (t, J=7.3 Hz, 3H)

Example 17:2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-2-(pyrrolidin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

3-Hydroxypicolinic acid (183 mg, 1.32 mmol) was dissolved in DMF (10 mL)and then2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(pyrrolidin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide.HCl(Intermediate P) (500 mg, 879 μmol), EDC.HCl (337 mg, 1.76 mmol), DIPEA(767 μL, 4.39 mmol) and HOBt (237 mg, 1.76 mmol) were added at 0° C. andstirred at RT for 16 hours. The RM was diluted with water and extractedwith 5% MeOH in DCM and washed with aq sat NaHCO₃ and brine and thecombined organic layers were dried over Na₂SO₄, filtered andconcentrated under reduced pressure. The crude product was purifiedtwice by column chromatography (2× Silica gel column: Silica 12 g,eluent DCM:MeOH 100:0 to 98:2). The obtained solid was stirred with 5%ACN and MeOH in Et₂O for 30 minutes, then sonicated for 10 minutes,filtered off, washed with n-pentane and dried to give the title compoundas an off-white solid.

LC-MS: Rt=1.04 min; MS m/z [M+H]⁺ 654.6, m/z [M−H]⁻ 652.4; UPLC-MS 1

LC-MS: Rt=5.08 min; MS m/z [M+H]⁺ 654.3, m/z [M−H]⁻ 652.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.37 (s, 1H), 9.99 (s, 1H), 8.06 (m, 1H),7.71 (d, J=8.4 Hz, 1H), 7.62 (m, 1H), 7.53 (d, J=8.4 Hz, 1H), 7.28 (m,2H), 5.14 (s, 2H), 4.54 (m, 1H), 3.49 (m, 3H), 3.37 (m, 4H), 3.20 (m,1H), 2.93 (m, 3H), 2.75 (m, 1H), 2.58 (m, 1H), 2.35 (s, 3H), 1.90 (m,4H), 1.16 (t, J=7.1 Hz, 3H)

Example 18:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-2-(pyrrolidin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To the stirred solution of 3-hydroxypicolinic acid (166 mg, 1.19 mmol),EDC.HCl (228 mg, 1.19 mmol), HOBt (161 mg, 1.19 mmol) in DMF (3 mL) wasaddedN-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(pyrrolidin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate Q) (330 mg, 596 μmol) and DIPEA (624 μL, 3.57 mmol) andthe RM was at RT for 16 hours. The RM was concentrated under reducedpressure and water was added. The resultant brown solid was filtered offand dried under vacuum. The crude product was purified by reverse phasepreparative HPLC (RP-HPLC acidic 4: 35 to 40% B in 2 min, 40 to 45% B in10 min) to give the title compound.

LC-MS: Rt=0.94 min; MS m/z [M+H]⁺ 675.3/677.3, m/z [M−H]⁻ 673.3/675.3;UPLC-MS 1

LC-MS: Rt=4.68 min; MS m/z [M+H]⁺ 675.2/677.2, m/z [M−H]⁻ 673.2/675.2;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.54 (s, br, 1H), 10.38 (s, br, 1H), 8.55(d, J=8.4 Hz, 1H), 8.06 (m, 1H), 7.95 (d, J=8.4 Hz, 1H), 7.28 (m, 2H),5.25 (s, 2H), 4.53 (m, 1H), 3.46 (m, 3H), 3.35 (m, 4H), 3.19 (m, 1H),2.91 (m, 3H), 2.75 (m, 1H), 2.57 (m, 1H), 1.89 (m, 4H), 1.15 (t, 3H)

Example 19:N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-2-(pyrrolidin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To the stirred solution ofN-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(pyrrolidin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide.HCl(Intermediate R) (150 mg, 249 μmol) in DCM (2 mL) was added3-hydroxypicolinoyl chloride (Intermediate CV) (43.0 mg, 273 μmol) at 0°C., followed by dropwise addition of DIPEA (217 μL, 1.24 mmol). The RMwas stirred at RT for 45 minutes. 3-Hydroxypicolinoyl chloride(Intermediate CV) (43.0 mg, 273 μmol) was added and the RM was stirredat RT for 12 hours. 3-Hydroxypicolinoyl chloride (Intermediate CV) (43.0mg, 273 μmol) was added and the RM was stirred at RT for 16 hours. TheRM was concentrated under reduced pressure. The crude product waspurified by reverse phase preparative HPLC (RP-HPLC neutral 3: 25 to 35%B in 2 min, 35 to 60% B in 8 min) to give the title compound.

LC-MS: Rt=1.11 min; MS m/z [M+H]⁺ 688.7/690.6, m/z [M−H]⁻ 686.4/688.3;UPLC-MS 1

LC-MS: Rt=5.52 min; MS m/z [M+H]⁺ 688.2/690.2, m/z [M−H]⁻ 686.3/688.3;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.37 (s, br, 1H), 10.08 (s, 1H), 8.06 (m,1H), 7.95 (s, 1H), 7.75 (s, 1H), 7.28 (m, 2H), 5.17 (s, 2H), 4.53 (m,1H), 3.46 (m, 3H), 3.36 (m, 4H), 3.20 (m, 1H), 2.93 (m, 3H), 2.75 (m,1H), 2.58 (m, 1H), 2.35 (s, 3H), 1.90 (m, 4H), 1.15 (t, J=7.3 Hz, 3H)

Example 20:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(4-((2,2-difluoroethyl)(methyl)amino)piperidin-1-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

ToN-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(4-((2,2-difluoroethyl)(methyl)amino)piperidin-1-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate S) (310 mg, 309 μmol) in DCM (3 mL) at 5° C. was added3-hydroxypicolinoyl chloride (Intermediate CV) (61.0 mg, 387 μmol) andthe RM was stirred for 3 minutes. DIPEA (162 μL, 928 μmol) was added andthe RM was allowed to warm to RT and stirred for 1 hour. Additional3-hydroxypicolinoyl chloride (Intermediate CV) (61.0 mg, 387 μmol) andDIPEA (162 μL, 928 μmol) were added and the RM was further stirred at RTfor 1 hour and 40 minutes. The RM was partitioned between DCM (20 mL)and 5% aq NaHCO₃ (20 mL). The organic layer was separated by filtrationthrough a phase separator. The aqueous layer was extracted with DCM (20mL). The organic layers were combined and evaporated in vacuo to give abrown gum. The crude product was purified by reverse phase preparativeHPLC (RP-HPLC acidic 1: 25 mL/min, 5 to 35% in 20 min with a plateau at35% for 1 min). The product containing fractions were combined andbasified with 5% aq NaHCO₃. Extracted with DCM (5×30 mL). The combinedorganic layers were filtered through a phase separator and evaporated invacuo to give an off-white foam. Further purification by columnchromatography (RediSep Column: Silica 4 g, eluent DCM:MeOH 100:0 to94:6) afforded a colourless solid which was recrystallized fromMeOH/water to give the title compound as a colourless powder.

LC-MS: Rt=0.73 min; MS m/z [M+H]⁺ 782.4/784.3, m/z [M−H]⁻ 780.5/782.5;UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.52 (s, 1H), 10.37 (s, 1H), 8.54 (d, J=8.2Hz, 1H), 8.07 (m, 1H), 7.96 (d, J=8.6 Hz, 1H), 7.29 (m, 2H), 6.15-5.85(m, br, 1H), 5.25 (s, 2H), 4.54 (m, 1H), 4.10 (m, 2H), 3.45 (m, 3H),3.22 (m, 1H), 2.94 (m, 3H), 2.78 (m, 4H), 2.59 (m, 1H), 2.27 (s, 3H),2.01 (m, 1H), 1.71 (m, 2H), 1.38 (m, 3H), 1.16 (t, J=7.4 Hz, 3H)

Example 21:2-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

3-Hydroxypicolinic acid (141 mg, 992 μmol) was dissolved in DCM (5.5 mL)at RT under argon. 1-chloro-N,N,2-trimethylprop-1-en-1-amine (149 mg,1.09 mmol) was added and the RM was stirred at RT for 1.25 hours. Asolution of2-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate U) (278 mg, 496 μmol) in DCM (2.5 mL) and DIPEA (260 μL,1.49 mmol) was added to the brown suspension. The resulting brownsolution was stirred at RT for 3.5 hours. The RM was quenched with water(5 mL) and aq sat NaHCO₃ (5 mL). It was extracted 4 times with DCM (4×40mL). The combined organic layers were washed twice with water, driedthrough a phase separator and concentrated under reduced pressure. Theresidue was adsorbed onto Isolute and purified by column chromatography(RediSep Column: Silica 24 g, eluent DCM:MeOH 100:0 to 90:10). Theproduct containing fractions were combined and concentrated. The solidwas submitted to SFC (SFC 5). The product containing fractions werecombined and concentrated to give the title compound as an off-beigesolid. A part of the solid was crystallized with MeOH (1.5 mL) and DCM(2 mL). The resulting solid was dried under HV to give the titlecompound.

LC-MS: Rt=0.97 min; MS m/z [M+H]⁺ 682.4, m/z [M−H]⁻ 680.3; UPLC-MS 3

¹H NMR (600 MHz, DMSO-d₆) δ 10.40 (s, 1H), 10.01 (s, 1H), 8.07 (m, 1H),7.72 (d, J=8.1 Hz, 1H), 7.63 (s, br, 1H), 7.54 (d, J=8.3 Hz, 1H), 7.29(m, 2H), 5.17 (s, 2H), 4.66 (m, 2H), 4.54 (m, 1H), 3.67 (d, J=12 Hz,2H), 3.58 (d, J=11.8 Hz, 2H), 3.49 (m, 2H), 3.40 (m, 1H), 3.21 (m, 1H),3.13 (m, 1H), 2.96 (m, 3H), 2.77 (m, 1H), 2.59 (m, 1H), 2.35 (s, 3H),1.89 (m, 1H), 1.15 (t, J=7.6 Hz, 3H)

Example 22:2-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-5-methyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide

1-Chloro-N,N,2-trimethylprop-1-en-1-amine (141 mg, 1.06 mmol) was addedto a solution of 3-hydroxypicolinic acid (134 mg, 961 μmol) in DCM (5mL) under argon and the RM was stirred at RT for 2 hours, then2-(2-(6-Oxa-3-azabicyclo[3.1.1]heptan-3-yl)-5-methyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide(Intermediate V) (441 mg, 481 μmol) dissolved in DCM (2.8 mL) was added,followed by DIPEA (420 μL, 2.40 mmol). The RM was stirred at RT for 2.2hours. 0.5 equivalent of the above described activated3-hydroxypicolinic acid solution was added again to the RM, followed byDIPEA (77.0 μL, 441 μmol). The RM was stirred at RT for 2 hours. The RMwas quenched with water (6 mL) and aq sat NaHCO₃ (6 mL) was added. Themixture was extracted with DCM (4×40 mL). The organic layer was washedwith aq sat NaHCO₃ and water, dried through a phase separator andconcentrated under reduced pressure. The crude product was adsorbed ontoIsolute and purified by column chromatography (RediSep Column: Silica 24g, eluent DCM:MeOH 100:0 to 90:10). The product containing fractionswere combined and concentrated under reduced pressure. The solid wasfurther purified by reverse phase preparative HPLC (RP-HPLC acidic 1: 15to 85% B in 20 min with a plateau at 85% for 1 min). The productcontaining fractions were combined and basified with a small amount ofaq sat NaHCO₃. The ACN was removed under reduced pressure and theresidue was extracted with DCM (3×40 mL). The combined organic layerswere washed with water (10 mL), then dried through a phase separator andconcentrated under reduced pressure to give the title compound as abeige solid.

LC-MS: Rt=0.93 min; MS m/z [M+H]⁺ 672.4, m/z [M−H]⁻ 670.4; UPLC-MS 3

¹H NMR (600 MHz, DMSO-d₆) δ 10.69 (s, br, 1H), 10.39 (s, br, 1H), 8.22(t, J=8.1 Hz, 1H), 8.06 (t, J=3.1 Hz, 1H), 7.80 (dd, J=2.1 Hz, 10.9 Hz,1H), 7.56 (dd, J=1.7 Hz, 8.4 Hz, 1H), 7.29 (m, 2H), 5.24 (s, 2H), 4.65(m, 2H), 4.52 (m, 1H), 3.66 (m, 2H), 3.57 (m, 2H), 3.46 (m, 2H), 3.38(m, 1H), 3.22 (m, 1H), 3.12 (m, 1H), 2.96 (m, 1H), 2.75 (m, 1H), 2.57(m, 1H), 2.47 (s, 3H), 1.89 (m, 1H)

Example 23:2-(2-((3R,4S)-3,4-difluoropyrrolidin-1-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

3-Hydroxypicolinic acid (152 mg, 1.07 mmol) was dissolved in DCM (6 mL)at RT under argon. 1-Chloro-N,N,2-trimethylprop-1-en-1-amine (161 mg,1.18 mmol) was added and the RM was stirred at RT for 1.2 hours. Asolution of2-(2-((3R,4S)-3,4-difluoropyrrolidin-1-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate W) (430 mg, 537 μmol) in DCM (3.5 mL) and DIPEA (281 μL,1.61 mmol) was added to the brown suspension. The resulting brownsolution was stirred at RT for 1 hour. The RM was quenched with water(10 mL), aq sat NaHCO₃ (5 mL) and extracted with DCM (4×40 mL). Thecombined organic layers were washed twice with water, dried through aphase separator and concentrated under reduced pressure. The residue wasadsorbed onto Isolute and purified by column chromatography (RediSepColumn: Silica 40 g, eluent DCM:MeOH 100:0 to 90:10). The productcontaining fractions were combined and concentrated under reducedpressure to give the title compound. A part of the solid wascrystallized from MeOH (1.5 mL) and DCM (2 mL). The resulting grey solidwas suspended in Et₂O and filtered, then dried under HV to give thetitle compound.

LC-MS: Rt=1.01 min; MS m/z [M+H]⁺ 690.3, m/z [M−H]⁻ 688.3; UPLC-MS 3

¹H NMR (600 MHz, DMSO-d₆) δ 10.39 (s, br, 1H), 10.01 (s, 1H), 8.06 (m,1H), 7.72 (d, J=8.3 Hz, 1H), 7.64 (s, br, 1H), 7.54 (d, J=8.3 Hz, 1H),7.29 (m, 2H), 5.45 (m, 1H), 5.36 (m, 1H), 5.16 (s, 2H), 4.54 (m, 1H),3.81 (m, 2H), 3.57 (m, 2H), 3.47 (m, 2H), 3.39 (m, 1H), 3.21 (m, 1H),2.95 (m, 3H), 2.76 (m, 1H), 2.58 (m, 1H), 2.36 (s, 3H), 1.16 (t, J=7.5Hz, 3H)

Example 24a:((R)-2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide)or((S)-2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide)and Example 24b:((R)-2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide)or((S)-2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide)

To the stirred solution ofN-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate Y) (300 mg, 504 μmol), 4-chloro-3-hydroxypicolinic acid(140 mg, 807 μmol), HOBt (136 mg, 1.01 mmol) and EDC.HCl (193 mg, 1.01mmol) in DCM (20 mL) was added pyridine (122 μL, 1.51 mmol) at 0° C. TheRM was stirred at RT for 16 hours. The RM was quenched with NaHCO₃ andextracted with DCM. The organic layer was dried over Na₂SO₄ andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (Silica gel column: Silica 4 g, eluent DCM:MeOH100:0 to 98:2). The residue was purified by preparative chiral HPLC(instrument: Agilent 1200 series, with single quad mass spectrometer;column: LUX CELLULOSE-4, 250 mm×21.1 mm, 5.0 μm; eluent: A=hexane,B=0.1% HCOOH in EtOH; flow rate: 15 mL/min; detection: 210 nm; injectionvolume: 0.9 mL; gradient: isocratic: 50(A):50(B)).

Example 24a: The product containing fractions were concentrated at 40°C. and washed with n-pentane (5×10 mL), decanted and dried to give thetitle compound as an off-white solid—first eluting stereoisomer.

Chiral HPLC (C-HPLC 2): Rt=10.764 min

LC-MS: Rt=1.08 min; MS m/z [M+H]⁺ 750.5/752.5, m/z [M−H]⁻ 748.4/750.4;UPLC-MS 1

LC-MS: Rt=5.29 min; MS m/z [M+H]⁺ 750.2/752.2, m/z [M−H]⁻ 748.2/750.2;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.68 (s, br, 2H), 8.56 (d, J=8.1 Hz, 1H),7.98 (d, J=5.6 Hz, 1H), 7.94 (d, J=8.1 Hz, 1H), 7.50 (d, J=5.1 Hz, 1H),6.72 (m, 1H), 5.34 (s, 2H), 4.53 (m, 1H), 3.52 (m, 4H), 3.28 (m, 4H),2.98 (m, 3H), 2.80 (m, 1H), 2.63 (m, 1H), 2.55 (m, 1H), 2.46 (m, 1H),2.16 (m, 2H), 1.95 (m, 1H), 1.68 (m, 1H), 1.17 (t, J=7.3 Hz, 3H)

Example 24b: The product containing fractions were concentrated at 40°C. and washed with n-pentane (5×10 mL), decanted and dried to give thetitle compound as an off-white solid—second eluting stereoisomer.

Chiral HPLC (C-HPLC 2): Rt=18.800 min

LC-MS: Rt=1.08 min; MS m/z [M+H]⁺ 750.1/752.1, m/z [M−H]⁻ 748.2/750.2;UPLC-MS 1

LC-MS: Rt=5.30 min; MS m/z [M+H]⁺ 750.1/752.1, m/z [M−H]⁻ 748.2/750.2;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.83 (s, br, 1H), 10.55 (s, br, 1H), 8.56(d, J=8.2 Hz, 1H), 8.06 (d, J=5.3 Hz, 1H), 7.92 (d, J=8.2 Hz, 1H), 7.55(d, J=5.3 Hz, 1H), 6.72 (m, 1H), 5.35 (s, 2H), 4.54 (m, 1H), 3.54 (m,4H), 3.28 (m, 3H), 3.25 (m, 1H), 2.99 (m, 3H), 2.81 (m, 1H), 2.62 (m,1H), 2.41 (m, 2H), 2.16 (m, 2H), 1.96 (m, 1H), 1.66 (m, 1H), 1.18 (t,J=7.3 Hz, 3H)

Example 25a:((R)—N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide)or((S)—N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide)and Example 25b:((R)—N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide)or((S)—N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide)

N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide.HCl(Intermediate Y) (120 mg, 190 μmol) and DIPEA (166 μL, 950 μmol) weredissolved in DCM (5 mL) and then 3-hydroxypicolinoyl chloride(Intermediate CV) (59.9 mg, 380 μmol) was added at 0° C. and stirred for2 hours. 3-hydroxypicolinoyl chloride (Intermediate CV) (59.9 mg, 380μmol) was added again and the reaction was continued under stirring for12 hours. The RM was diluted with DCM and washed with water and aqNaHCO₃ (2×20 mL), washed with water and brine, dried over Na₂SO₄,filtered and concentrated. The crude product was combined with anotherexperiment and purified by column chromatography (Silica gel column:Silica 4 g, eluent DCM:MeOH 100:0 to 99:1) then further purified byreverse phase preparative HPLC (RP-HPLC acidic 10: 40 to 50% B in 2 min,50 to 60% B in 8 min) to give the title compound as an off-white solid.

The racemate was purified by preparative chiral HPLC (instrument:Agilent 1200 series, with single quad mass spectrometer; column:CELLULOSE-4, 250 mm×21.2 mm; eluent: A=hexane, B=0.1% HCOOH in MeOH:EtOH1:1; flow rate: 20 mL/min; detection: 210 nm; injection volume: 0.9 mL;gradient: isocratic 60(A):40(B)).

Example 25a: First eluting stereoisomer, off-white solid.

Chiral HPLC (C-HPLC 1): Rt=10.070 min

LC-MS: Rt=0.98 min; MS m/z [M+H]⁺ 716.5/718.6, m/z [M−H]⁻ 714.3/716.3;UPLC-MS 1

LC-MS: Rt=4.76 min; MS m/z [M+H]⁺ 716.2/718.2, m/z [M−H]⁻ 714.2/716.2;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.46 (s, br, 2H), 8.56 (d, J=8.5 Hz, 1H),8.05 (m, 1H), 7.90 (d, J=8.4 Hz, 1H), 7.28 (m, 2H), 6.72 (m, 1H), 5.30(s, 2H), 4.54 (m, 1H), 3.47 (m, 4H), 3.27 (s, 3H), 3.21 (m, 1H), 2.96(m, 3H), 2.79 (m, 1H), 2.59 (m, 3H), 2.43 (m, 1H), 2.14 (m, 1H), 1.95(m, 1H), 1.67 (m, 1H), 1.17 (t, J=7.2 Hz, 3H)

Example 25b: Second eluting stereoisomer, off-white solid.

Chiral HPLC (C-HPLC 1): Rt=16.023 min

LC-MS: Rt=0.96 min; MS m/z [M+H]⁺ 716.3/718.3, m/z [M−H]⁻ 714.3/716.3;UPLC-MS 1

LC-MS: Rt=4.77 min; MS m/z [M+H]⁺ 716.2/718.2, m/z [M−H]⁻ 714.2/716.2;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.39 (s, br, 2H), 8.56 (d, J=8.0 Hz, 1H),8.06 (m, 1H), 7.93 (d, J=8.1 Hz, 1H), 7.28 (m, 2H), 6.72 (m, 1H), 5.32(s, 2H), 4.54 (m, 1H), 3.46 (m, 4H), 3.27 (s, 3H), 3.20 (m, 1H), 2.96(m, 3H), 2.79 (m, 1H), 2.59 (m, 3H), 2.41 (m, 1H), 2.14 (m, 1H), 1.95(m, 1H), 1.68 (m, 1H), 1.17 (t, J=7.1 Hz, 3H)

Example 26:rac-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

4-Chloro-3-hydroxypicolinic acid (118 mg, 851 μmol) was dissolved in DMF(5 mL) and thenrac-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide.HCl(Intermediate Z) (260 mg, 425 μmol), EDC.HCl (163 mg, 851 μmol), DIPEA(372 μL, 2.13 mmol) and HOBt (115 mg, 851 μmol) were added at 0° C. andthe RM was stirred at RT for 14 hours. The RM was diluted with water andextracted with 5% MeOH in DCM and washed with aq sat NaHCO₃, brine andthe combined organic layers were dried over Na₂SO₄, filtered andconcentrated under reduced pressure. The crude product was purified byreverse phase preparative HPLC (RP-HPLC acidic 5: 30 to 40% B in 2 min,40 to 50% B in 8 min) to give the title compound.

LC-MS: Rt=0.90 min; MS m/z [M+H]⁺ 696.3, m/z [M−H]⁻ 694.3; UPLC-MS 1

LC-MS: Rt=4.38 min; MS m/z [M+H]⁺ 696.3, m/z [M−H]⁻ 694.4; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.26 (m, 2H), 8.19 (d, J=8.3 Hz, 1H), 8.06(m, 1H), 7.72 (d, J=8.2 Hz, 1H), 7.28 (m, 2H), 6.72 (m, 1H), 5.27 (s,2H), 4.54 (m, 1H), 3.47 (m, 4H), 3.28 (m, 4H), 2.98 (m, 3H), 2.80 (m,1H), 2.57 (m, 6H), 2.15 (m, 1H), 1.96 (m, 1H), 1.68 (m, 1H), 1.19 (t,J=7.5 Hz, 3H)

Example 27:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AA) (492 mg, 692 μmol) was dissolved in DCM (7 mL) at 0°C. under argon. 3-Hydroxypicolinoyl chloride (Intermediate CV) (163 mg,1.04 mmol) was added to the suspension, followed by slow addition ofDIPEA (483 μL, 2.77 mmol) and the solution was stirred at RT for 1.5hours. 3-Hydroxypicolinoyl chloride (Intermediate CV) (25.0 mg, 159μmol) and DIPEA (320 μL, 1.84 mmol) were added again and the RM wascontinued stirring at RT for 2.3 hours. The reaction was quenched byaddition of water (5 mL) and aq sat NaHCO₃ (5 mL). Then it was extracted4 times with DCM (4×40 mL). The organic layer was washed with water (5mL), aq sat NaHCO₃ (5 mL) and water again (10 mL). The organic layer wasdried through a phase separator and concentrated under reduced pressure.The residue was adsorbed onto Isolute and purified by columnchromatography (RediSep Column: Silica 40 g Gold, eluent DCM:MeOH 100:0to 90:10). The product containing fractions were combined andconcentrated under reduced pressure. The resulting solid was purified byreverse phase preparative HPLC (RP-HPLC acidic 1: 15 to 85% in 20 minwith a plateau at 85% for 1 min). The product containing fractions werecombined, basified with aq sat NaHCO₃ (5 mL). The ACN was removed underreduced pressure. The aqueous layer was washed 4 times with DCM (4×35mL). The combined organic layers were washed with water (10 mL), driedthrough a phase separator and concentrated under reduced pressure togive the title compound as a white solid.

LC-MS: Rt=0.87 min; MS m/z [M+H]⁺ 690.3/692.3, m/z [M−H]⁻ 688.1/690.1;UPLC-MS 1

Example 28:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(5-methoxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To the stirred solution of 5-methoxy-6-methylpyrimidine-4-carboxylicacid (Intermediate CW) (133 mg, 794 μmol) in DCM (9 mL) at 0° C. wereadded EDC.HCl (203 mg, 1.06 mmol), pyridine (128 μL, 1.59 mmol) and HOBt(143 mg, 1.06 mmol) and the RM was stirred at 0° C. for 10 minutes, thenwas addedN-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AC) (300 mg, 529 μmol) and the RM was stirred at RT for 16hours. The RM was diluted with DCM and washed with aq sat NaHCO₃, washedwith water and the combined organic layers were dried over Na₂SO₄,concentrated and dried. The crude product was purified by columnchromatography (Silica gel column: Silica 12 g, eluent DCM:MeOH 100:0 to99:1) to give the title compound.

LC-MS: Rt=1.51 min; MS m/z [M+H]⁺ 717.2/719.2; UPLC-MS 11

Step 2:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To the stirred solution ofN-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(5-methoxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(250 mg, 310 μmol) in DMF (3 mL) was added LiCl (132 mg, 3.10 mmol) andthe RM was heated at 150° C. for 3 hours. LiCl (132 mg, 3.10 mmol) wasadded again and the RM was heated at 150° C. for 4 hours. The RM wasquenched with water and extracted with 10% MeOH in DCM (3×50 mL) anddried over Na₂SO₄, filtered, concentrated and dried. The crude productwas purified by column chromatography (Silica gel column: Silica 12 g,eluent DCM:MeOH 100:0 to 98:2). The residue was purified by reversephase preparative HPLC (RP-HPLC acidic 4: 15 to 25% B in 2 min, 25 to55% B in 7 min) and the product containing fractions were concentratedbelow 40° C. and dried to give the title compound as an off-white solid.

LC-MS: Rt=0.97 min; MS m/z [M+H]⁺ 703.2/705.2, m/z [M−H]⁻ 701.3/703.2;UPLC-MS 1

LC-MS: Rt=4.73 min; MS m/z [M+H]⁺ 703.2/705.1, m/z [M−H]⁻ 701.3/703.2;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.52 (s, br, 1H), 10.25 (s, br, 1H), 8.57(m, 2H), 7.95 (d, J=8.3 Hz, 1H), 5.86 (m, 1H), 5.35 (s, 2H), 4.52 (m,1H), 4.10 (m, 2H), 3.49 (m, 3H), 3.25 (m, 1H), 2.99 (m, 3H), 2.81 (m,1H), 2.64 (m, 1H), 2.44 (s, 3H), 2.17 (m, 2H), 1.84 (m, 2H), 1.18 (t,J=7.6 Hz, 3H)

Example 29:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AC) (700 mg, 1.24 mmol) was suspended in DMF (12 mL),perfluorophenyl 3-hydroxypicolinate (Intermediate CT) (754 mg, 2.47mmol) and Et₃N (342 μL, 2.47 mmol) were added at RT and the RM wasstirred at 80° C. for 16 hours. The RM was extracted three times withDCM. The combined organic layers were washed with brine, dried overNa₂SO₄ and concentrated under reduced pressure. The crude product waspurified by column chromatography (Silica gel column: Silica 12 g,eluent hexane:EtOAc 100:0 to 60:40). The product containing fractionswere concentrated and dried under HV to give the title compound.

LC-MS: Rt=0.94 min; MS m/z [M+H]⁺ 688.5/690.5, m/z [M−H]⁻ 686.2/688.2;UPLC-MS 1

LC-MS: Rt=4.58 min; MS m/z [M+H]⁺ 688.2/690.2, m/z [M−H]⁻ 686.2/688.2;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.54 (s, 1H), 10.38 (s, 1H), 8.57 (d, J=8.5Hz, 1H), 8.06 (m, 1H), 7.95 (d, J=8.5 Hz, 1H), 7.28 (m, 2H), 5.85 (m,1H), 5.35 (s, 2H), 4.54 (m, 1H), 4.10 (m, 2H), 3.42 (m, 3H), 3.21 (m,1H), 2.96 (m, 3H), 2.79 (m, 1H), 2.61 (m, 1H), 2.16 (m, 2H), 1.84 (m,2H), 1.17 (t, J=7.3 Hz, 3H)

Example 30:2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide

To the stirred solution of 4-chloro-3-hydroxypicolinic acid (282 mg,1.62 mmol) in DMF (5 mL) were added DIPEA (354 μL, 2.03 mmol) and PyAOP(635 mg, 1.22 mmol) at 0° C. After 10 minutes was addedN-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AC) (460 mg, 811 μmol) to the RM at 0° C. The RM wasstirred at RT for 16 hours. Water was added and the mixture wasextracted with EtOAc. The organic layer was dried over Na₂SO₄ andconcentrated under reduced pressure. The crude product was purified byreverse phase preparative HPLC (RP-HPLC acidic 5: 20 to 30% B in 2 min,30 to 60% B in 8 min) to give the title compound.

LC-MS: Rt=1.04 min; MS m/z [M+H]⁺ 722.5/724.5/726.5, m/z [M−H]⁻720.3/722.2/724.2; UPLC-MS 1

LC-MS: Rt=5.07 min; MS m/z [M+H]⁺ 722.2/724.1/726.2, m/z [M−H]⁻720.2/722.2/724.2; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.83 (s, 1H), 10.55 (s, 1H), 8.58 (d, J=8.4Hz, 1H), 8.07 (d, J=5.0 Hz, 1H), 7.96 (d, J=8.4 Hz, 1H), 7.56 (d, J=5.0Hz, 1H), 5.86 (m, 1H), 5.36 (s, 2H), 4.54 (m, 1H), 4.10 (m, 2H), 3.54(m, 3H), 3.24 (m, 1H), 2.98 (m, 3H), 2.81 (m, 1H), 2.65 (m, 1H), 2.16(m, 2H), 1.84 (m, 2H), 1.18 (t, J=7.1 Hz, 3H)

Example 31:2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

Step 1:2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(5-methoxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

To the stirred solution of2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide(Intermediate AD) (600 mg, 1.10 mmol),5-methoxy-6-methylpyrimidine-4-carboxylic acid (Intermediate CW) (222mg, 1.32 mmol) and HATU (626 mg, 1.65 mmol) in DMF (15 mL) was addedDIPEA (288 μL, 1.65 mmol) at 0° C. The RM was stirred at RT for 16hours. The RM was concentrated under reduced pressure to give the titlecompound.

LC-MS: Rt=1.48 min; MS m/z [M+H]⁺ 697.3; UPLC-MS 11

Step 2:2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

To the stirred solution of2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(5-methoxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide(250 mg, 255 μmol) in DMF (2 mL) was added LiCl (108 mg, 2.55 mmol) andthe RM was stirred at 200° C. for 1 hour in the MW. The RM was quenchedwith water and extracted with 10% MeOH in DCM (3×50 mL) and dried overNa₂SO₄, filtered, concentrated and dried. The crude product was purifiedby column chromatography (Silica gel column: Silica 12 g, eluentDCM:MeOH 100:0 to 98:2). The residue after the column chromatography waspurified by reverse phase preparative HPLC (RP-HPLC acidic 10: 15 to 25%B in 2 min, 15 to 60% B in 10 min). The product containing fractionswere concentrated to get 45 mg solid, which was combined with anotherbatch, washed with 30% Et₂O in n-hexane, decanted and dried to give thetitle compound as an off-white solid.

LC-MS: Rt=0.90 min; MS m/z [M+H]⁺ 683.6, m/z [M−H]⁻ 681.4; UPLC-MS 1

LC-MS: Rt=4.34 min; MS m/z [M+H]⁺ 683.3, m/z [M−H]⁻ 681.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.22 (s, br, 2H), 8.57 (s, 1H), 8.20 (d,J=8.0 Hz, 1H), 7.73 (d, J=8.2 Hz, 1H), 5.86 (m, 1H), 5.27 (s, 2H), 4.52(m, 1H), 4.10 (m, 2H), 3.50 (m, 3H), 3.27 (m, 1H), 2.99 (m, 3H), 2.81(m, 1H), 2.65 (m, 1H), 2.58 (s, 3H), 2.44 (s, 3H), 2.18 (m, 2H), 1.85(m, 2H), 1.19 (t, J=7.2 Hz, 3H)

Example 32:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AE) (590 mg with 80% purity, 833 μmol) was dissolved inDCM (10 mL) and 3-hydroxypicolinoyl chloride (Intermediate CV) (197 mg,1.25 mmol) was added, followed by DIPEA (437 μL, 2.50 mmol). The RM wasstirred at RT for 1.5 hours. 3-Hydroxypicolinoyl chloride (IntermediateCV) (36.0 mg, 228 μmol) was added. The RM was stirred at RT for 1 hour.DIPEA (1.00 mL, 5.73 mmol) was added. The RM was stirred at RT for 2hours. 3-Hydroxypicolinoyl chloride (Intermediate CV) (116 mg, 736 μmol)was added. The RM was stirred at RT for 1 hour. Water (10 mL), aq satNaHCO₃ (10 mL) and DCM (10 mL) were added. The aqueous layer was washedtwice with DCM (2×10 mL). The combined organic layers were dried througha phase separator and concentrated under reduced pressure. The crudeproduct was purified in 5 portions by reverse phase preparative HPLC(5×RP-HPLC acidic 1: 5 to 100% B). All product containing fractionspurer than 95% were combined, basified with aq sat NaHCO₃, extractedtwice with DCM (2×15 mL), dried through a phase separator andconcentrated under reduced pressure. The concentrated fractions weresuspended in MeOH and sonicated for 1 minute. Then it was filtered, thecake was washed with MeOH (500 μL) and dried under HV to give the titlecompound. All product containing fractions with the impurity at Rt=1.05min were combined, basified with aq sat NaHCO₃, extracted twice with DCM(2×15 mL), dried through a phase separator and concentrated underreduced pressure. All product containing fractions with the impurity atRt=1.13 min were combined, basified with aq sat NaHCO₃, extracted twicewith DCM (2×15 mL), dried through a phase separator and concentratedunder reduced pressure. Both impure fractions were combined andsuspended in MeOH (10 mL). Then it was sonicated for 30 minutes andfiltered. The cake was washed with a small amount of MeOH (10 mL) anddried under HV to give 90% pure product. The cake was suspended in MeOH(10 mL) and ACN (10 mL) and stirred at 40° C. for 2 hours. It wasfiltered, the cake was washed with MeOH (1 mL) and dried under HV togive the title compound.

Both pure fractions were dissolved in DCM (10 mL) and EtOH (10 mL) andleft at RT for 5 days. The solid was filtered off and washed with asmall amount of Et₂O. The cake was dried under HV to give the titlecompound.

LC-MS: Rt=1.08 min; MS m/z [M+H]⁺ 687.2/689.2, m/z [M−H]⁻ 685.4/687.5;UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.38 (s, 1H), 10.33 (s, 1H), 8.07 (m, 2H),7.96 (s, 1H), 7.71 (d, J=8.6 Hz, 1H), 7.29 (m, 2H), 5.87 (m, 1H), 5.31(s, 2H), 4.55 (m, 1H), 4.10 (m, 2H), 3.43 (m, 3H), 3.23 (m, 1H), 2.97(m, 3H), 2.80 (m, 1H), 2.62 (m, 1H), 2.17 (m, 2H), 1.84 (m, 2H), 1.18(t, J=7.6 Hz, 3H)

Example 33:N-(4-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

N-(4-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediates AG) (340 mg, 600 μmol) was suspended in DMF (10 mL) andperfluorophenyl 3-hydroxypicolinate (Intermediate CT) (366 mg, 1.20mmol) and Et₃N (166 μL, 1.20 mmol) were added and the RM was stirred at70° C. for 3 hours. The crude product was purified by reverse phasepreparative HPLC (RP-HPLC acidic 3: 10 to 20% B in 2 min, 20 to 60% B in10 min) to give the title compound.

LC-MS: Rt=0.84 min; MS m/z [M+H]⁺ 688.3/690.3, m/z [M−H]⁻ 686.3/688.3;UPLC-MS 1

LC-MS: Rt=4.15 min; MS m/z [M+H]⁺ 688.2/690.2, m/z [M−H]⁻ 686.2/688.2;UPLC-MS 2

¹H NMR (600 MHz, DMSO-d₆) δ 10.65 (s, 1H), 10.36 (s, 1H), 9.06 (s, 1H),8.22 (s, 1H), 8.02 (m, 1H), 7.25 (m, 2H), 6.72 (m, 1H), 5.29 (s, 2H),4.50 (m, 1H), 4.21 (m, 2H), 3.76 (m, 2H), 3.42 (m, 2H), 3.35 (m, 1H),3.18 (m, 1H), 2.93 (m, 3H), 2.76 (m, 1H), 2.58 (m, 1H), 2.52 (m, 2H),1.15 (t, J=7.6 Hz, 3H)

Example 34:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide(Intermediate AH) (450 mg, 823 μmol) was suspended in DMF (5 mL).Perfluorophenyl 3-hydroxypicolinate (Intermediate CT) (503 mg, 1.65mmol) and Et₃N (228 μL, 1.65 mmol) were added and the RM was stirred at70° C. for 3 hours. The RM was concentrated under reduced pressure. Thecrude product was purified by reverse phase preparative HPLC (RP-HPLCacidic 11: 30 to 40% B in 2 min, 40 to 70% B in 10 min) to give thetitle compound.

LC-MS: Rt=0.77 min; MS m/z [M+H]⁺ 668.3, m/z [M−H]⁻ 666.3; UPLC-MS 1

LC-MS: Rt=3.79 min; MS m/z [M+H]⁺ 668.3, m/z [M−H]⁻ 666.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.36 (m, br, 2H), 8.78 (s, 1H), 8.05 (m,1H), 7.85 (s, 1H), 7.28 (m, 2H), 6.83 (m, 1H), 5.25 (s, 2H), 4.55 (m,1H), 4.26 (m, 2H), 3.81 (m, 2H), 3.45 (m, 3H), 3.22 (m, 1H), 3.00 (m,3H), 2.80 (m, 1H), 2.62 (m, 1H), 2.50 (m, 2H), 2.38 (s, 3H), 1.20 (t,J=7.3 Hz, 3H)

Example 35:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

To a solution of2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide.TFA(Intermediate AI) (400 mg, 606 μmol) in DMF (4 mL) was added Et₃N (252μL, 1.82 mmol), followed by dropwise addition of perfluorophenyl3-hydroxypicolinate (Intermediate CT) (185 mg, 606 μmol) at 0° C. Thenthe RM was allowed to warm to RT and it was stirred at 80° C. for 14hours. The crude product was concentrated under reduced pressure,extracted with water and purified by column chromatography (Silica gelcolumn: Silica 12 g, eluent hexane:EtOAc 100:0 to 20:80). The productwas recrystallized with ACN to give the title compound.

LC-MS: Rt=0.81 min; MS m/z [M+H]⁺ 668.6, m/z [M−H]⁻ 666.4; UPLC-MS 1

LC-MS: Rt=3.91 min; MS m/z [M+H]⁺ 668.3, m/z [M−H]⁻ 666.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.38 (s, 1H), 10.23 (s, 1H), 8.19 (d, J=8.4Hz, 1H), 8.06 (m, 1H), 7.73 (d, J=8.4 Hz, 1H), 7.28 (m, 2H), 6.82 (m,1H), 5.28 (s, 2H), 4.55 (m, 1H), 4.26 (m, 2H), 3.80 (m, 2H), 3.44 (m,3H), 3.22 (m, 1H), 2.97 (m, 3H), 2.80 (m, 1H), 2.62 (m, 1H), 2.57 (s,3H), 2.52 (m, 2H), 1.19 (t, J=7.2 Hz, 3H)

Example 36:2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

To a solution of 4-chloro-3-hydroxypicolinic acid (423 mg, 2.44 mmol) inDCM (10 mL) were added DIPEA (500 μL, 2.86 mmol) and PyAOP (829 mg, 1.59mmol). After the colour changed to dark brown, the RM was stirred for 10minutes and then was added dropwise a solution of2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide.TFA(Intermediate AI) (700 mg, 1.06 mmol) in DCM (10 mL) and DIPEA (500 μL,2.86 mmol). The RM was stirred at RT for 18 hours. The RM was pouredinto aq sat NaHCO₃ and extracted several times with EtOAc. The combinedorganic layers were dried through a phase separator and concentrated.The crude product was purified by column chromatography (Silica gelcolumn: Silica 12 g, eluent hexane:EtOAc 100:0 to 0:100). The productcontaining fractions were combined and concentrated under reducedpressure. Then washed with Et₂O. The solid was purified again by columnchromatography (Silica gel column: Silica 12 g, eluent hexane:EtOAc100:0 to 5:95) to give the title compound.

LC-MS: Rt=0.91 min; MS m/z [M+H]⁺ 702.3/704.3, m/z [M−H]⁻ 700.3/702.3;UPLC-MS 1

LC-MS: Rt=4.49 min; MS m/z [M+H]⁺ 702.2/704.2, m/z [M−H]⁻ 700.3/702.3;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.83 (s, br, 1H), 10.22 (s, br, 1H), 8.19(d, J=8.8 Hz, 1H), 8.06 (d, J=5.0 Hz, 1H), 7.73 (d, J=8.4 Hz, 1H), 7.55(d, J=5.0 Hz, 1H), 6.82 (m, 1H), 5.28 (s, 2H), 4.54 (m, 1H), 4.26 (m,2H), 3.80 (m, 2H), 3.50 (m, 3H), 3.15 (m, 1H), 3.00 (m, 3H), 2.81 (m,1H), 2.65 (m, 1H), 2.57 (s, 3H), 2.48 (m, 2H), 1.19 (t, J=7.0 Hz, 3H)

Example 37:2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

4-Chloro-3-hydroxypicolinic acid (233 mg, 1.34 mmol) was suspended inDCM (7 mL) and 1-chloro-N,N,2-trimethylprop-1-en-1-amine (190 μL, 1.44mmol) was added. After 5 minutes most of the solid was dissolved. Thelight suspension was stirred at RT for 2.5 hours. The RM was cooled to0° C. and2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AJ) (431 mg, 765 μmol) was added, followed by DIPEA (601μL, 3.44 mmol). The yellow solution turned into a black solution. The RMwas stirred at RT for 20 minutes. Water (10 mL), aq sat NaHCO₃ (10 mL)and DCM (10 mL) were added. The aqueous layer was washed twice with DCM(2×10 mL). The combined organic layers were dried through a phaseseparator and concentrated under reduced pressure and concentrated underreduced pressure. The crude product was purified by columnchromatography (RediSep Column: Silica 40 g, eluent DCM:DCM/MeOH (8/2)100:0 to 50:50). The product containing fractions were combined,concentrated under vacuum and dried under HV to afford a brown solid.Then it was purified in three portions by reverse phase preparative HPLC(RP-HPLC acidic 1: 20 to 75% B in 20 min with a plateau at 75% for 1min, RP-HPLC acidic 1: 25 to 80% B in 20 min with a plateau at 80% for 1min and RP-HPLC acidic 1: 40 to 80% B in 20 min with a plateau at 80%for 1 min). The product containing fractions were combined, basifiedwith aq sat NaHCO₃, extracted twice with DCM (2×15 mL), dried through aphase separator and concentrated under reduced pressure. All impurefractions were combined, basified with aq sat NaHCO₃, extracted twicewith DCM, dried through a phase separator and concentrated under reducedpressure. Then it was suspended in ACN (2 mL) and sonicated for 2minutes, filtered, combined with the pure fractions and concentratedunder reduced pressure to give the title compound. The solid wasdissolved in EtOH (10 mL) and DCM (15 mL), filtered and left standing atRT for 3 days to crystallize. Then it was filtered and washed with Et₂O.The cake was dried under HV to give the title compound.

The sodium salt was prepared analogous to the general procedure.

LC-MS: Rt=1.11 min; MS m/z [M+H]⁺ 719.4/721.4, m/z [M−H]⁻ 717.5/719.5;UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.83 (s, 1H), 10.20 (s, 1H), 8.06 (d, J=5.1Hz, 1H), 7.57 (m, 3H), 6.82 (m, 1H), 5.26 (s, 2H), 4.54 (m, 1H), 4.26(m, 2H), 3.81 (m, 2H), 3.55 (m, 3H), 3.25 (m, 1H), 3.00 (m, 3H), 2.81(m, 1H), 2.65 (m, 1H), 2.50 (m, 2H), 2.24 (s, 3H), 1.19 (t, J=7.4 Hz,3H)

Example 38:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AJ) (504 mg, 894 μmol) and 3-hydroxypicolinoyl chloride(Intermediate CV) (254 mg, 1.61 mmol) were mixed in DCM (7 mL) and DIPEA(312 μL, 1.79 mmol) were added. The dark solution was stirred at RT for1 hour. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL) were added.The aqueous layer was washed twice with DCM (2×10 mL). The combinedorganic layers were dried through a phase separator and concentratedunder reduced pressure and concentrated under reduced pressure. Thecrude product was purified by column chromatography (RediSep Column:Silica 24 g, eluent DCM:DCM/MeOH (8/2) 100:0 to 35:65). The productcontaining fractions were combined, concentrated under vacuum and driedunder HV to afford a white solid. The solid was dissolved in EtOH (5 mL)and DCM (15 mL) and left standing until it crystallized out to give thetitle compound.

LC-MS: Rt=1.02 min; MS m/z [M+H]⁺ 685.3, m/z [M−H]⁻ 683.4; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.38 (s, 1H), 10.20 (s, 1H), 8.06 (m, 1H),7.58 (m, 2H), 7.29 (m, 2H), 6.82 (m, 1H), 5.25 (s, 2H), 4.55 (m, 1H),4.26 (m, 2H), 3.81 (m, 2H), 3.44 (m, 3H), 3.22 (m, 1H), 2.98 (m, 3H),2.80 (m, 1H), 2.62 (m, 1H), 2.52 (m, 2H), 2.24 (s, 3H), 1.19 (t, J=7.3Hz, 3H)

Example 39:2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

The reaction was performed in 4 batches obtained as follows.4-Chloro-3-hydroxypicolinic acid (400 mg, 2.31 mmol) was dissolved inDCM (36 mL) at RT under argon. 1-chloro-N,N,2-trimethylprop-1-en-1-amine(360 mg, 2.66 mmol) was added and the RM was stirred at RT for 2.5hours. At 0° C. were addedN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AK) (1.10 g, 1.58 mmol) and DIPEA (1.23 mL, 7.08 mmol).The resulting brown solution was stirred at RT for 1.3 hours. Thecombined RM from the 4 batches was quenched with water (40 mL) and aqsat NaHCO₃ (40 mL). It was extracted 4 times with DCM (4×200 mL). Thecombined organic layers were washed with water (50 mL) and brine (50mL), dried through a phase separator and concentrated under reducedpressure. The residue was adsorbed onto Isolute and purified in 2portions by column chromatography (RediSep Column: Silica 120 g Gold,eluent DCM:DCM/MeOH (1/1) 100:0 to 60:40) and (RediSep Column: Silica120 g Gold, eluent DCM:DCM/MeOH (1/1) 100:0 to 90:10). The productcontaining fractions were combined and concentrated under reducedpressure to give the title compound as a beige solid.

LC-MS: Rt=1.13 min; MS m/z [M+H]⁺ 721.4/723.4/725.4, m/z [M−H]⁻719.5/721.5/723.5; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.83 (s, 1H), 10.35 (s, 1H), 8.06 (m, 2H),7.96 (s, 1H), 7.71 (dd, J=2.1 Hz, 8.7 Hz, 1H), 7.55 (d, J=5.1 Hz, 1H),6.83 (m, 1H), 5.32 (s, 2H), 4.54 (m, 1H), 4.25 (m, 2H), 3.80 (m, 2H),3.53 (m, 3H), 3.26 (m, 1H), 2.99 (m, 3H), 2.82 (m, 1H), 2.65 (m, 1H),2.52 (m, 2H), 1.19 (t, J=7.1 Hz, 3H)

Example 40:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-fluoro-3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-(4-(3-(benzyloxy)-4-fluoropicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

To a stirred solution ofN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide.TFA(Intermediate AK) (300 mg, 441 μmol), 3-(benzyloxy)-4-fluoropicolinicacid (Intermediate CU) (115 mg, 463 μmol) and HATU (201 mg, 529 μmol) inDMF (10 mL) was added DIPEA (385 μL, 2.21 mmol) at RT and the RM wasstirred at RT for 5 minutes. The RM was diluted with EtOAc/water,extracted twice with EtOAc and the combined organic extracts were driedover Na₂SO₄ and concentrated. The crude product was purified by columnchromatography (RediSep Column: Silica 24 g, eluent DCM:DCM/MeOH (8/2)100:0 to 50:50). The product containing fractions were combined andconcentrated to afford a white foam.

LC-MS: Rt=1.23 min; MS m/z [M+H]⁺ 795.3/797.3, m/z [M−H]⁻ 793.4/795.4;UPLC-MS 1

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-fluoro-3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To a stirred suspension of2-(6-(4-(3-(benzyloxy)-4-fluoropicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(289 mg, 363 μmol) in DCM (10 mL) was added boron trichloride methylsulfide complex (363 μL, 727 μmol) at RT and the RM was stirred at RTfor 20 hours. The RM was quenched with MeOH. It was diluted inDCM/NaHCO₃, extracted twice with DCM and the combined organic extractswere washed with water and brine, dried over Na₂SO₄ and concentrated.The crude product was purified by column chromatography (RediSep Column:Silica 24 g, eluent DCM:DCM/MeOH (8/2) 100:0 to 60:40). The productcontaining fractions were combined and concentrated to give an off-whitesolid. This solid was dissolved in EtOH (6 mL) at 50° C. and the RM wasleft at RT for 4 hours. A white solid was filtered to give the titlecompound.

The sodium salt was prepared analogous to the general procedure.

LC-MS: Rt=1.03 min; MS m/z [M+H]⁺ 705.4/707.4, m/z [M−H]⁻ 703.5/705.5;UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.80 (s, br, 1H), 10.36 (s, 1H), 8.06 (m,2H), 7.96 (m, 1H), 7.71 (dd, J=2.1 Hz, 8.8 Hz, 1H), 7.34 (dd, J=5.3 Hz,10.9 Hz, 1H), 6.83 (m, 1H), 5.31 (s, 2H), 4.54 (m, 1H), 4.25 (m, 2H),3.80 (m, 2H), 3.46 (m, 3H), 3.23 (m, 1H), 2.98 (m, 3H), 2.80 (m, 1H),2.63 (m, 1H), 2.51 (m, 2H), 1.18 (t, J=7.5 Hz, 3H)

Example 41:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-fluoro-3-hydroxy-6-methylpicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-(4-(3-(benzyloxy)-4-fluoro-6-methylpicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AK) (227 mg, 334 μmol),3-(benzyloxy)-4-fluoro-6-methylpicolinic acid (Intermediate CX) (110 mg,371 μmol) and HATU (140 mg, 367 μmol) were suspended in DCM (5 mL) andcooled to 0° C. Then DIPEA (204 μL, 1.17 mmol) was added and the RM wasstirred at RT for 2.5 hours. Water (10 mL), aq sat NaHCO₃ (10 mL) andDCM (10 mL) were added. The aqueous layer was washed twice with DCM(2×10 mL). The combined organic layers were tried through a phaseseparator and concentrated under reduced pressure. The crude product waspurified by reverse phase preparative HPLC (RP-HPLC basic 1: 5 to 95% Bin 20 min with a plateau at 95% for 1 min). The product containingfractions were combined, basified with aq sat NaHCO₃, extracted twicewith DCM, dried through a phase separator and concentrated under reducedpressure to give the title compound.

LC-MS: Rt=1.27 min; MS m/z [M+H]⁺ 809.5/811.5, m/z [M−H]⁻ 807.2/809.2;UPLC-MS 1

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-fluoro-3-hydroxy-6-methylpicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

2-(6-(4-(3-(Benzyloxy)-4-fluoro-6-methylpicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(242 mg, 292 μmol) was dissolved in DCM (5 mL) and TFA (5.00 mL, 64.9mmol) was added and the RM was stirred at 60° C. overnight. After 1night ca 50% conversion. The RM was continued stirring at 60° C. for onefurther night. The RM was concentrated under reduced pressure. The crudeproduct was purified by reverse phase preparative ISCO (RediSep Column:C18 50 g Gold, eluent water+0.1% TFA:ACN 100:0 to 0:100). The productcontaining fractions were combined, basified with aq sat NaHCO₃,extracted twice with DCM, dried through a phase separator andconcentrated under reduced pressure to give the title compound.

LC-MS: Rt=1.06 min; MS m/z [M+H]⁺ 719.5/721.6, m/z [M−H]⁻ 717.4/719.4;UPLC-MS 1

LC-MS: Rt=5.31 min; MS m/z [M+H]⁺ 719.5/721.5, m/z [M−H]⁻ 717.5/719.3;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.35 (s, 2H), 8.05 (d, J=8.3 Hz, 1H), 7.96(d, J=2.1 Hz, 1H), 7.71 (dd, J=2.1 Hz, 8.7 Hz, 1H), 7.22 (d, J=11.8 Hz,1H), 6.83 (m, 1H), 5.31 (s, 2H), 4.52 (m, 1H), 4.25 (m, 2H), 3.80 (m,2H), 3.47 (m, 3H), 3.22 (m, 1H), 2.98 (m, 3H), 2.80 (m, 1H), 2.64 (m,1H), 2.51 (m, 2H), 2.39 (s, 3H), 1.18 (t, J=7.1 Hz, 3H)

Example 42:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

To a stirred solution ofN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AK) (300 mg, 429 μmol),5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (Intermediate CY)(115 mg, 472 μmol) and HATU (245 mg, 644 μmol) in DMF (3 mL) was addedDIPEA (375 μL, 2.15 mmol) at RT and the RM was stirred at RT for 15minutes. The RM was diluted with EtOAc/water, extracted twice with EtOAcand the combined organic extracts were dried over Na₂SO₄ andconcentrated. The crude product was purified by column chromatography(RediSep Column: Silica 24 g, eluent DCM:MeOH 100:0 to 90:10). Theproduct containing fractions were combined and concentrated to give thetitle compound as a beige foam. The product was dissolved in EtOH,stirred at 60° C. over 18 hours, then cooled down to 0° C., filtered offand washed with EtOH to give the title compound as a white solid.

LC-MS: Rt=1.20 min; MS m/z [M+H]⁺ 792.4/794.4, m/z [M−H]⁻ 790.6/792.6;UPLC-MS 1

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To a stirred suspension of2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(266 mg, 312 μmol) in DCM (6 mL) was added boron trichloride methylsulfide complex (312 μL, 625 μmol) at RT and the RM was stirred at RTfor 14 hours. The RM was quenched with MeOH. Then it was diluted withDCM/water, extracted twice with DCM and the combined organic extractswere washed with water and brine, dried over Na₂SO₄ and concentrated.The crude product was purified by column chromatography (RediSep Column:Silica 24 g, eluent DCM:MeOH 100:0 to 93:07). The product containingfractions were combined and concentrated to give the title compound asan off-white solid. The product was dissolved in EtOH, stirred at 60° C.over 18 h, then cooled down to 0° C., filtered off and washed with EtOHto give the title compound as a white solid characterized by the XRPDdiffractogram in FIG. 1 . The table Ex 42a below shows the mostprominent peaks (deg 2theta) of the XRPD diffractogram of FIG. 1 . TheXRPD was repeated on a title compound sample as prepared by theprocedure described in Ex 42 above, then purified by columnchromatography (RediSep 150 g, eluent DCM:MeOH 100:0 to 90:10) followedby trituration in EtOH. The sample was characterized by the XPRDdiffractogram in FIG. 7 . The table Ex 42b below shows the mostprominent peaks (deg 2theta) of the XRPD diffractogram of FIG. 7 .

LC-MS: Rt=1.05 min; MS m/z [M+H]⁺ 702.4/704.4, m/z [M−H]⁻ 700.5/702.5;UPLC-MS 1

¹H NMR (600 MHz, DMSO-d₆) δ 10.40 (s, 1H), 10.20 (br s, 1H), 8.57 (s,1H), 8.06 (d, J=8.5 Hz, 1H), 7.97 (d, J=2.0 Hz, 1H), 7.72 (dd, J=8.7 Hz,J=2.1 Hz, 1H), 6.83 (m, 1H), 5.32 (m, 2H), 4.52 (m, 1H), 4.25 (m, 2H),3.80 (t, J=5.5 Hz, 2H), 3.50 (m, 3H), 3.25 (m, 1H), 2.99 (m, 3H), 2.81(m, 1H), 2.63 (m, 1H), 2.51 (m, 2H), 2.44 (s, 3H), 1.18 (t, J=7.5 Hz,3H).

The sodium salt was prepared analogous to the general procedure:

¹H NMR (400 MHz, DMSO-d₆) δ 10.36 (s, br, 2H), 8.55 (s, 1H), 8.05 (d,J=8.5 Hz, 1H), 7.96 (m, 1H), 7.71 (dd, J=2.1 Hz, 8.8 Hz, 1H), 6.83 (m,1H), 5.32 (s, 2H), 4.52 (m, 1H), 4.25 (m, 2H), 3.80 (m, 2H), 3.48 (m,3H), 3.25 (m, 1H), 2.99 (m, 3H), 2.81 (m, 1H), 2.64 (m, 1H), 2.52 (m,2H), 2.43 (s, 3H), 1.18 (t, J=7.4 Hz, 3H).

TABLE Ex 42a Angle d Value 2-Theta° Angstrom Intensity 11.85 7.46 medium13.71 6.45 medium 14.46 6.12 low 15.33 5.78 medium 17.03 5.20 medium18.33 4.84 high 19.98 4.44 medium 22.42 3.96 medium 22.95 3.87 medium27.20 3.28 low

TABLE Ex42b Angle d Value 2-Theta° Angstrom Intensity 6.78 13.033 high8.97 9.846 low 11.88 7.446 medium 13.55 6.530 low 13.74 6.441 low 14.486.112 medium 15.34 5.771 high 16.83 5.264 low 17.03 5.203 low 18.304.843 high 19.49 4.550 low 19.94 4.449 low 21.28 4.172 low 21.51 4.127low 22.38 3.970 medium 22.91 3.879 medium 23.27 3.819 low 25.41 3.502low 27.26 3.269 medium 29.03 3.074 low 29.78 2.998 low 29.96 2.980 low

Example 43:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide-2,2-d2

A beige suspension ofN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Example 42) (70.0 mg, 100 μmol) and K2CO₃ (13.8 mg, 100 μmol) in THF(997 μL)/DMSO-d6 (282 μL, 3.99 mmol)/D2O (180 μL, 9.97 mmol) was heatedup to 65° C. and stirred for 45 minutes. A small amount of water wasadded to the RM, which was then extracted 3 times with EtOAc. Thecombined organic layers were washed once with brine, dried over Na₂SO₄,concentrated and dried under reduced pressure to give a beige solid. Theaqueous layer showed still product in it. The aqueous layer wasconcentrated to remove THF, then it was extracted twice with DCM. Theaqueous layer was diluted with aq sat NaHCO₃ then extracted three timeswith DCM. The combined organic layers were dried through a phaseseparator, concentrated and dried under vacuum to give the titlecompound.

LC-MS: Rt=1.04 min; MS m/z [M+H]⁺ 704.5/706.5, m/z [M−H]⁻ 702.2/704.2;UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.36 (s, 2H), 8.56 (s, 1H), 8.06 (d, J=8.4Hz, 1H), 7.96 (d, J=2.1 Hz, 1H), 7.71 (dd, J=2.1 Hz, 8.8 Hz, 1H), 6.83(m, 1H), 4.52 (m, 1H), 4.25 (m, 2H), 3.80 (m, 2H), 3.49 (m, 3H), 3.25(m, 1H), 2.98 (m, 3H), 2.81 (m, 1H), 2.64 (m, 1H), 2.52 (m, 2H), 2.44(s, 3H), 1.19 (t, J=7.2 Hz, 3H)

Example 44:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxypyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxypyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AK) (508 mg, 727 μmol), 5-methoxypyrimidine-4-carboxylicacid (134 mg, 872 μmol) and HATU (427 mg, 1.09 mmol) were mixed in DMF(10 mL) at RT under argon. DIPEA (635 μL, 3.64 mmol) was added and theRM was stirred at RT for 2 hours. The reaction was quenched with water(10 mL). Aq sat NaHCO₃ (15 mL) was added and the mixture was extractedwith EtOAc (3×70 mL). The organic layer was washed with water (20 mL)and brine (20 mL), dried over Na₂SO₄, filtered through a phase separatorand concentrated under reduced pressure. The residue was adsorbed ontoIsolute and purified by column chromatography (RediSep Column: Silica 40g, eluent DCM:DCM/MeOH (8/2) 100:0 to 50:50). The product containingfractions were combined, concentrated, mixed with Et₂O, sonicated,filtered and dried under HV to give the title compound as a beige solid.

LC-MS: Rt=0.99 min; MS m/z [M+H]⁺ 702.2/704.2, m/z [M−H]⁻ 700.4/702.4;UPLC-MS 1

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxypyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxypyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(178 mg, 254 μmol) was mixed with DMF (2 mL) and LiCl (43.0 mg, 1.01mmol) was added and the RM was stirred at 150° C. for 18.3 hours. Thenit was allowed to cool to RT. The reaction was diluted with water and aqsat NaHCO₃ (5 mL). It was extracted with EtOAc (3×20 mL). The organiclayer was washed with water (5 mL) and brine (2×5 mL). The organic layerwas washed again with EtOAc (3×20 mL). The combined organic layers weredried over NaSO₄, filtered through a phase separator and concentratedunder reduced pressure. The residue was adsorbed onto Isolute andpurified by column chromatography (RediSep Column: Silica 40 g, eluentDCM:DCM/MeOH (8/2) 100:0 to 35:65). The product containing fractionswere combined and concentrated under reduced pressure. The solid waspurified by reverse phase preparative HPLC (RP-HPLC basic 1: 15 to 65% Bin 20 min with a plateau at 65% for 1 min). The product containingfractions were combined, the ACN was removed under reduced pressure andthe residue was lyophilized to give the title compound as a beige solid.

LC-MS: Rt=0.98 min; MS m/z [M+H]⁺ 688.3/690.3, m/z [M−H]⁻ 686.3/688.3;UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.75 (s, br, 1H), 10.35 (s, br, 1H), 8.70(s, 1H), 8.44 (s, 1H), 8.05 (d, J=8.5 Hz, 1H), 7.96 (d, J=2.2 Hz, 1H),7.71 (dd, J=2.1 Hz, 8.9 Hz, 1H), 6.83 (m, 1H), 5.31 (s, 2H), 4.51 (m,1H), 4.25 (m, 2H), 3.80 (m, 2H), 3.47 (m, 3H), 3.23 (m, 1H), 2.97 (m,3H), 2.81 (m, 1H), 2.63 (m, 1H), 2.50 (m, 2H), 1.18 (t, J=7.4 Hz, 3H)

Example 45:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AL) (212 mg, 337 μmol) and 3-hydroxypicolinoyl chloride(Intermediate CV) (89.0 mg, 565 μmol) were dissolved in DCM (5 mL) andDIPEA (294 μL, 1.68 mmol) was added. The RM was stirred at RT for 30minutes. 3-Hydroxypicolinoyl chloride (Intermediate CV) (89.0 mg, 565μmol) was added again and the RM was continued stirring at RT for 1hour. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL) were added.The aqueous layer was washed twice with DCM (2×10 mL). The combinedorganic layers were dried through a phase separator and concentratedunder reduced pressure. The crude product was purified by columnchromatography (RediSep Column: Silica 40 g, eluent DCM:DCM/MeOH (9/1)100:0 to 60:40). The product containing fractions were combined,concentrated under vacuum and dried under HV to afford a beige solid.The crude product was purified by reverse phase preparative HPLC(RP-HPLC acidic 1: 20 to 80% B in 20 min with a plateau at 80% for 1min). The product containing fractions were combined, basified with aqsat NaHCO₃, extracted twice with DCM (2×15 mL), dried through a phaseseparator and concentrated under reduced pressure. The product wasdissolved in DCM (5 mL) and MeOH (2 mL) and left to stand at RTovernight. The resulting crystals were filtered off and washed with Et₂O(4×3 mL). The cake was dried under HV to give the title compound aswhite crystals.

LC-MS: Rt=0.92 min; MS m/z [M+H]⁺ 688.4/690.4, m/z [M−H]⁻ 686.4/688.3;UPLC-MS 3

¹H NMR (400 MHz, DMSO-d₆) δ 10.56 (s, 1H), 10.38 (s, 1H), 8.56 (d, J=8.0Hz, 1H), 8.07 (m, 1H), 7.96 (d, J=8.0 Hz, 1H), 7.29 (m, 2H), 6.82 (m,1H), 5.36 (s, 2H), 4.55 (m, 1H), 4.25 (m, 2H), 3.80 (m, 2H), 3.46 (m,3H), 3.23 (m, 1H), 2.98 (m, 3H), 2.80 (m, 1H), 2.62 (m, 1H), 2.51 (m,2H), 1.18 (t, J=7.3 Hz, 3H)

Example 46:2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide

4-Chloro-3-hydroxypicolinic acid (1.48 g, 8.54 mmol) was suspended inDCM (140 mL) and 1-chloro-N,N,2-trimethylprop-1-en-1-amine (1.29 mL,9.73 mmol) was added. The red-brown solution turned into a yellowsolution after 1 hour. The RM was stirred at RT for 2.5 hours.N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AL) (5.31 g, 8.43 mmol) and DIPEA (3.90 mL, 22.3 mmol)were dissolved in DCM (80 mL) and the prepared acid chloride solutionwas added dropwise during 1 hour. 5% aq NaHCO₃ and DCM were added. Theaqueous layer was washed twice with DCM (2×100 mL). The combined organiclayers were dried through a phase separator and concentrated underreduced pressure. The crude product was purified by columnchromatography (RediSep Column: Silica 330 g, eluent DCM:MeOH 100:0 to90:10). The product containing fractions were combined, concentrated anddried under HV. The residue was purified by reverse phase preparativeISCO (RediSep Column: C18 240 g, eluent water:ACN 100:0 to 0:100). Theproduct containing fractions were combined, concentrated and dried underreduced pressure. The resulting solid was suspended at 55° C. in EtOH(25 mL), stirred overnight at 55° C., cooled to RT, stirred at 0° C. for1 hour, filtered off, and dried under HV to give the title compound.

LC-MS: Rt=0.99 min; MS m/z [M+H]⁺ 722.3/724.3/726.3, m/z [M−H]⁻720.2/722.2/724.2; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.79 (s, br, 1H), 10.59 (s, br, 1H), 8.57(d, J=8.1 Hz, 1H), 8.05 (d, J=4.9 Hz, 1H), 7.95 (d, J=8.5 Hz, 1H), 7.54(d, J=4.9 Hz, 1H), 6.82 (m, 1H), 5.36 (s, 2H), 4.54 (m, 1H), 4.25 (m,2H), 3.80 (m, 2H), 3.53 (m, 3H), 3.23 (m, 1H), 2.99 (m, 3H), 2.81 (m,1H), 2.66 (m, 1H), 2.51 (m, 2H), 1.18 (t, J=7.4 Hz, 3H)

Example 47:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To the stirred solution of 5-methoxy-6-methylpyrimidine-4-carboxylicacid (Intermediate CW) (311 mg, 1.85 mmol) in DMF (10 mL) at RT wereadded EDC.HCl (355 mg, 1.85 mmol), HOBt (250 mg, 1.85 mmol) and pyridine(300 μL, 3.71 mmol). ThenN-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AL) (700 mg, 1.24 mmol) was added and the RM was stirredat RT for 14 hours. Water (20 mL) was added to the RM and it wasextracted with 10% MeOH in DCM (2×20 mL). The organic layer was washedwith brine (20 mL), dried over Na₂SO₄, filtered and concentrated. Thecrude product was purified by column chromatography (Silica gel column:Silica 12 g, eluent DCM:MeOH 100:0 to 95:5) to give the title compound.The product was heated in EtOH:DCM (1:2) until fully dissolved. Thesolution was evaporated at RT until dry, then dried under vacuum to givethe title compound as beige solid.

LC-MS: Rt=1.48 min; MS m/z [M+H]⁺ 717.1/719.1; UPLC-MS 11

Step 2:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To the stirred solution ofN-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(700 mg, 820 μmol) in DMF (10 mL) at RT, was added LiCl (348 mg, 8.20mmol) and the RM was stirred at 140° C. for 14 hours. Water (20 mL) wasadded to the RM and it was extracted twice with DCM (2×20 mL). Theorganic layer was washed with brine (20 mL), dried over Na₂SO₄, filteredand concentrated. The crude product was purified by reverse phasepreparative HPLC (RP-HPLC acidic 4: 30 to 40% B in 2 min, 40 to 55% B in8 min) to give the title compound. The product was heated in 1:2EtOH:DCM until fully dissolved. The solution was evaporated at RT untildry, then dried under vacuum to give the title compound as beige solidcharacterized by the XRPD diffractogram in FIG. 3 . The table belowshows the most prominent peaks (deg 2theta) of the XRPD diffractogram ofFIG. 3 .

The sodium salt was prepared analogous to the general procedure.

LC-MS: Rt=0.92 min; MS m/z [M+H]⁺ 703.4/705.4, m/z [M−H]⁻ 701.5/703.5;UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.46 (s, br, 2H), 8.56 (m, 2H), 7.95 (d,J=8.4 Hz, 1H), 6.82 (m, 1H), 5.35 (s, 2H), 4.52 (m, 1H), 4.25 (m, 2H),3.80 (m, 2H), 3.48 (m, 3H), 3.25 (m, 1H), 2.99 (m, 3H), 2.81 (m, 1H),2.65 (m, 1H), 2.51 (m, 2H), 2.44 (s, 3H), 1.18 (t, J=7.7 Hz, 3H)

Angle d Value 2-Theta° Angstrom Intensity 9.45 9.35 medium 12.75 6.94high 13.28 6.66 medium 21.69 4.09 medium 25.25 3.52 high 26.85 3.32medium

Example 48:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-fluoro-3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-(4-(3-(benzyloxy)-4-fluoropicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide

To a stirred suspension ofN-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AL) (445 mg, 785 μmol), 3-(benzyloxy)-4-fluoropicolinicacid (Intermediate CU) (204 mg, 824 μmol) and HATU (358 mg, 942 μmol) inDMF (10 mL) was added DIPEA (685 μL, 3.92 mmol) at RT and the RM wasstirred at RT for 5 minutes. The RM was diluted in EtOAc/water,extracted twice with EtOAc and the combined organic extracts were washedwith water and brine, dried over Na₂SO₄ and concentrated. The crudeproduct was purified by column chromatography (RediSep Column: Silica 24g, eluent DCM:DCM/MeOH (8/2) 100:0 to 50:50). The product containingfractions were combined and concentrated to give the title compound as ayellow foam.

LC-MS: Rt=1.14 min; MS m/z [M+H]⁺ 796.5/798.5, m/z [M−H]⁻ 794.5/796.5;UPLC-MS 1

Step 2:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-fluoro-3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To a stirred suspension of2-(6-(4-(3-(benzyloxy)-4-fluoropicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide(610 mg, 758 μmol) in DCM (1 mL) was added TFA (5.00 mL, 64.9 mmol) at50° C. and the RM was stirred at 50° C. for 3 hours, then at 45° C. for18 hours. The RM was diluted with DCM/NaHCO₃, extracted twice with DCMand the combined organic extracts were washed with brine, dried overNa₂SO₄ and concentrated. The crude product was purified by columnchromatography (RediSep Column: Silica 40 g, eluent DCM:DCM/MeOH (8/2)100:0 to 60:40). The product containing fractions were combined andconcentrated. This solid was dissolved at 50° C. in EtOH (20 mL) andleft at RT overnight. A white solid was filtered off to give the titlecompound as a white solid.

LC-MS: Rt=0.91 min; MS m/z [M+H]⁺ 706.2/708.1, m/z [M−H]⁻ 704.5/706.5;UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.65 (s, br, 2H), 8.56 (d, J=8.3 Hz, 1H),8.07 (m, 1H), 7.95 (d, J=8.4 Hz, 1H), 7.35 (m, 1H), 6.82 (m, 1H), 5.35(s, 2H), 4.54 (m, 1H), 4.25 (m, 2H), 3.80 (m, 2H), 3.45 (m, 3H), 3.23(m, 1H), 2.97 (m, 3H), 2.80 (m, 1H), 2.62 (m, 1H), 2.50 (m, 2H), 1.18(t, J=7.3 Hz, 3H)

Example 49:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-fluoro-3-hydroxy-6-methylpicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-(4-(3-(benzyloxy)-4-fluoro-6-methylpicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide

N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AL) (277 mg, 407 μmol),3-(benzyloxy)-4-fluoro-6-methylpicolinic acid (Intermediate CX) (133 mg,447 μmol) and HATU (170 mg, 447 μmol) were suspended in DCM (5 mL) andcooled to 0° C. Then DIPEA (249 μL, 1.42 mmol) was added and the RM wasstirred at RT for 2 hours. Water (20 mL), aq sat NaHCO₃ (10 mL) and DCM(30 mL) were added. The aqueous layer was washed twice with DCM (2×20mL). The combined organic layers were dried through a phase separatorand concentrated under reduced pressure. The crude product was purifiedby reverse phase preparative ISCO (RediSep Column: C18 26 g, eluentwater+0.1% TFA:ACN 90:10 to 0:100). The product containing fractionswere combined, basified with aq sat NaHCO₃, extracted twice with DCM(2×15 mL), dried through a phase separator and concentrated underreduced pressure to give the title compound.

LC-MS: Rt=1.17 min; MS m/z [M+H]⁺ 810.5/812.5, m/z [M−H]⁻ 808.2/810.2;UPLC-MS 1

Step 2:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-fluoro-3-hydroxy-6-methylpicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

2-(6-(4-(3-(Benzyloxy)-4-fluoro-6-methylpicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide(244 mg, 289 μmol) was dissolved in DCM (5 mL) and TFA (5.00 mL, 64.9mmol) was added. The RM was stirred at 50° C. for 2 days. The RM wasconcentrated under reduced pressure. The crude product was purified byreverse phase preparative ISCO (RediSep Column: C18 50 g Gold, eluentwater+0.1% TFA:ACN 90:10 to 0:100). The product containing fractionswere combined, basified with aq sat NaHCO₃, extracted twice with DCM(2×15 mL), dried through a phase separator and concentrated underreduced pressure to give the title compound.

LC-MS: Rt=0.93 min; MS m/z [M+H]⁺ 720.6/722.5, m/z [M−H]⁻ 718.4/720.3;UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.55 (s, 1H), 10.35 (s, 1H), 8.56 (d, J=7.9Hz, 1H), 7.95 (d, J=8.5 Hz, 1H), 7.22 (d, J=11.6 Hz, 1H), 6.82 (m, 1H),5.36 (s, 2H), 4.52 (m, 1H), 4.25 (m, 2H), 3.80 (m, 2H), 3.47 (m, 3H),3.23 (m, 1H), 2.98 (m, 3H), 2.80 (m, 1H), 2.63 (m, 1H), 2.50 (m, 2H),2.39 (s, 3H), 1.18 (t, J=7.2 Hz, 3H)

Example 50:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

Step 1:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

2-(2-(3,6-Dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AM) (700 mg, 1.24 mmol) was suspended in DMF (5 mL) and5-methoxy-6-methylpyrimidine-4-carboxylic acid (Intermediate CW) (292mg, 1.74 mmol) was added at 0° C. PyAOP (971 mg, 1.86 mmol) and DIPEA(651 μL, 3.73 mmol) were added and the RM was stirred at RT for 12hours. The RM was concentrated under reduced pressure. Water was addedand the mixture was filtered to give the title compound.

LC-MS: Rt=1.64 min; MS m/z [M+H]⁺ 714.1; UPLC-MS 12

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

2-(2-(3,6-Dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide(800 mg, 953 μmol) was suspended in DMF (5 mL) and LiCl (182 mg, 4.29mmol) was added. The reaction was stirred in the MW at 200° C. for 20minutes. The reaction was concentrated under reduced pressure. The crudeproduct was purified by reverse phase preparative HPLC (RP-HPLC acidic2: 40 to 50% B in 2 min, 50 to 60% B in 9 min) to give the titlecompound.

The sodium salt was prepared analogous to the general procedure.

LC-MS: Rt=1.02 min; MS m/z [M+H]⁺ 700.5, m/z [M−H]⁻ 698.4; UPLC-MS 1

LC-MS: Rt=5.10 min; MS m/z [M+H]⁺ 700.3, m/z [M−H]⁻ 698.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.22 (s, br, 1H), 10.06 (s, 1H), 8.57 (s,1H), 7.78 (d, J=13 Hz, 1H), 7.67 (d, J=8.3 Hz, 1H), 6.81 (m, 1H), 5.29(s, 2H), 4.52 (m, 1H), 4.25 (m, 2H), 3.80 (m, 2H), 3.49 (m, 3H), 3.27(m, 1H), 2.99 (m, 3H), 2.82 (m, 1H), 2.65 (m, 1H), 2.52 (m, 2H), 2.44(s, 3H), 2.35 (s, 3H), 1.19 (t, J=7.4 Hz, 3H)

Example 51:N-(2-chloro-4-(pentafluorosulfanyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(pentafluorosulfanyl)phenyl)acetamide

N-(2-chloro-4-(pentafluorosulfanyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AN) (296 mg, 360 μmol),5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (Intermediate CY)(97.0 mg, 396 μmol) and HATU (144 mg, 378 μmol) were mixed in DCM (5 mL)and DIPEA (189 μL, 1.08 mmol) was added. The suspension turned into asolution and was stirred at RT for 2 hours. Water (10 mL), aq sat NaHCO₃(10 mL) and DCM (10 mL) were added. The aqueous layer was washed twicewith DCM (2×10 mL). The combined organic layers were dried through aphase separator and concentrated under reduced pressure. The crudeproduct was purified by reverse phase preparative HPLC (RP-HPLC acidic1: 10 to 90% B in 20 min with a plateau at 90% for 1 min). The productcontaining fractions were combined, basified with aq sat NaHCO₃,extracted twice with DCM, dried through a phase separator andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (RediSep Column: Silica 40 g, eluent DCM:DCM/MeOH(1/1) 100:0 to 0:100). The product containing fractions were combined,concentrated under vacuum and dried under HV to give the title compound.

LC-MS: Rt=1.25 min; MS m/z [M+H]⁺ 850.5/852.5, m/z [M−H]⁻ 848.5/850.5;UPLC-MS 1

Step 2:N-(2-chloro-4-(pentafluorosulfanyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(pentafluorosulfanyl)phenyl)acetamide(195 mg, 227 μmol) was dissolved in DCM (3 mL) and TFA (3.00 mL, 38.9mmol) was added and the RM was stirred at 70° C. for 1 day. The RM wasconcentrated under reduced pressure. Water (10 mL), aq sat NaHCO₃ (10mL) and DCM (10 mL) were added. The aqueous layer was washed twice withDCM (2×10 mL). The combined organic layers were dried through a phaseseparator and concentrated under reduced pressure. The crude product waspurified by column chromatography (RediSep Column: Silica 24 g, eluentDCM:DCM/MeOH (1/1) 100:0 to 90:10). The product containing fractionswere combined and concentrated under reduced pressure. A part of thesolid was dissolved in DCM:ACN (8:2), filtered and left standing at RT.The resulting crystals were filtered off, suspended in Et₂O, washed witha small amount of Et₂O and dried under HV to give the title compound.

LC-MS: Rt=1.12 min; MS m/z [M+H]⁺ 760.4/762.4, m/z [M−H]⁻ 758.4/760.4;UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.40 (s, 1H), 10.22 (s, 1H), 8.58 (s, 1H),8.15 (d, J=2.7 Hz, 1H), 8.09 (d, J=9.3 Hz, 1H), 7.90 (dd, J=2.5 Hz, 9.2Hz, 1H), 6.82 (m, 1H), 5.33 (s, 2H), 4.52 (m, 1H), 4.25 (m, 2H), 3.80(m, 2H), 3.50 (m, 3H), 3.25 (m, 1H), 2.98 (m, 3H), 2.81 (m, 1H), 2.66(m, 1H), 2.50 (m, 2H), 2.44 (s, 3H), 1.18 (t, J=7.4 Hz, 3H)

Example 52:N-(2-chloro-4-(pentafluorosulfanyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxypyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:N-(2-chloro-4-(pentafluorosulfanyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxypyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

N-(2-chloro-4-(pentafluorosulfanyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AN) (256 mg, 312 μmol), 5-methoxypyrimidine-4-carboxylicacid (67.9 mg, 441 μmol) and HATU (160 mg, 421 μmol) were mixed in DCM(5 mL) and DIPEA (210 μL, 1.20 mmol) was added. The suspension turnedinto a solution and was stirred at RT for 2 hours. Then it was stored inthe fridge overnight. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10mL) were added. The aqueous layer was washed twice with DCM (2×10 mL).The combined organic layers were dried through a phase separator andconcentrated under reduced pressure. The crude product was purified in 2portions by reverse phase preparative HPLC (RP-HPLC acidic 1: 5 to 95% Bin 20 min with a plateau at 95% for 1 min and RP-HPLC acidic 1: 15 to80% B in 20 min with a plateau at 80% for 1 min). The product containingfractions were combined, basified with aq sat NaHCO₃, extracted twicewith DCM, dried through a phase separator and concentrated under reducedpressure. It was purified by column chromatography (RediSep Column:Silica 40 g, eluent DCM:DCM/MeOH (1/1) 100:0 to 90:10). The productcontaining fractions were combined, concentrated under vacuum and driedunder HV to afford the title compound.

LC-MS: Rt=1.04 min; MS m/z [M+H]⁺ 760.0/762.0, m/z [M−H]⁻ 758.4/760.4;UPLC-MS 1

Step 2:N-(2-chloro-4-(pentafluorosulfanyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxypyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

N-(2-chloro-4-(pentafluorosulfanyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxypyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(169 mg, 222 μmol) and LiCl (37.7 mg, 889 μmol) were mixed with DMF (2mL) and stirred at 140° C. under sealed conditions for 1 day. Water (10mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL) were added. The aqueous layerwas washed twice with DCM (2×10 mL). The combined organic layers weredried through a phase separator and concentrated under reduced pressure.The crude product was purified by reverse phase preparative HPLC(RP-HPLC acidic 1: 10 to 90% B in 20 min with a plateau at 90% for 1min). The product containing fractions were combined, basified with aqsat NaHCO₃, extracted twice with DCM, dried through a phase separatorand concentrated under reduced pressure. It was purified by columnchromatography (RediSep Column: Silica 24 g, eluent DCM:DCM/MeOH (1/1)100:0 to 75:25). The product containing fractions were combined,concentrated under vacuum and dried under HV to afford the titlecompound as a pale yellow solid.

LC-MS: Rt=1.02 min; MS m/z [M+H]⁺ 746.4/748.4, m/z [M−H]⁻ 744.4/746.3;UPLC-MS 1

LC-MS: Rt=5.20 min; MS m/z [M+H]⁺ 746.1/748.1, m/z [M−H]⁻ 744.2/746.2;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, br, 1H), 10.40 (s, br, 1H), 8.70(s, 1H), 8.44 (s, 1H), 8.15 (d, J=2.6 Hz, 1H), 8.09 (d, J=9.2 Hz, 1H),7.90 (dd, J=2.6 Hz, 9.2 Hz, 1H), 6.82 (m, 1H), 5.32 (s, 2H), 4.51 (m,1H), 4.25 (m, 2H), 3.80 (m, 2H), 3.47 (m, 3H), 3.27 (m, 1H), 2.98 (m,3H), 2.81 (m, 1H), 2.65 (m, 1H), 2.52 (m, 2H), 1.18 (t, J=7.6 Hz, 3H)

Example 53:N-(4-(difluoromethyl)-5-fluoro-2-methylphenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(difluoromethyl)-5-fluoro-2-methylphenyl)acetamide

ToN-(4-(difluoromethyl)-5-fluoro-2-methylphenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AO) (880 mg, 1.61 mmol) and5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (Intermediate CY)(394 mg, 1.61 mmol) in DMF (12 mL) was added DIPEA (1.41 mL, 8.07 mmol)followed by HATU (675 mg, 1.77 mmol). The RM was stirred at RT for 15minutes. The RM was diluted with water (50 mL) and EtOAc (50 mL) and theresulting precipitate was stirred at RT overnight. The brown solid wasfiltered and washed with water (10 mL). The solid was redissolved in DCM(70 mL), dried over Na₂SO₄, filtered and evaporated in vacuo to give thetitle compound as a brown solid—contains aldehyde side product

LC-MS: Rt=1.08 min; MS m/z [M+H]⁺ 772.6, m/z [M−H]⁻ 770.4; UPLC-MS 1

Step 2:N-(4-(difluoromethyl)-5-fluoro-2-methylphenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

2-(6-(4-(5-(Benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(difluoromethyl)-5-fluoro-2-methylphenyl)acetamide(1.10 g, 1.43 mmol) was heated in TFA (5.00 mL, 64.9 mmol) at 50° C. for1 hour. LCMS indicated minor peak for desired product at Rt=0.90 min[M+H]⁺ 682/683. Significant hydrolysis of CF₂H to aldehyde. The RM wasevaporated in vacuo to give a brown oil. The crude product wasredissolved in MeOH and purified in 3 portions by reverse phasepreparative HPLC (RP-HPLC acidic 1: 20 to 48% B in 20 min with a plateauat 48% for 1 min and 2×RP-HPLC acidic 1: 25 to 46% B in 20 min with aplateau at 46% for 1 min). The impure fractions containing >2% aldehydeside-product were combined and the pH basified by addition of aq satNaHCO₃, extracted with DCM (2×50 mL). The organic layers were combined,dried over Na₂SO₄, filtered and evaporated to give a pale brown solid.The brown solid was redissolved in MeOH and purified by reverse phasepreparative HPLC (RP-HPLC basic 1: 20 to 50% B in 10 min with a plateauat 50% for 1 min). The pure product containing fractions were extractedwith DCM (2×50 mL). The organic layers were combined, dried over Na₂SO₄,filtered and evaporated to give the title compound as a colourlesssolid.

LC-MS: Rt=0.89 min; MS m/z [M+H]⁺ 682.2, m/z [M−H]⁻ 680.4; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.22 (s, br, 1H), 9.99 (s, 1H), 8.57 (s,1H), 7.61 (d, J=12.2 Hz, 1H), 7.50 (d, J=8.1 Hz, 1H), 7.26-7.00 (t, br,J=54.9 Hz, 1H), 6.82 (m, 1H), 5.26 (s, 2H), 4.52 (m, 1H), 4.26 (m, 2H),3.80 (m, 2H), 3.48 (m, 3H), 3.26 (m, 1H), 3.01 (m, 3H), 2.81 (m, 1H),2.64 (m, 1H), 2.50 (m, 2H), 2.44 (s, 3H), 2.31 (s, 3H), 1.19 (t, J=7.1Hz, 3H)

Example 54:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-4-formyl-2-methylphenyl)acetamide

Side product of Example 53.

The fractions from the acidic RP Prep HPLC containing the aldehydeside-product >97% pure were combined and the pH basified by addition ofaq sat NaHCO₃, then it was extracted with DCM (2×50 mL). The organiclayers were combined, dried over Na₂SO₄, filtered and evaporated to givethe title compound as a beige solid.

LC-MS: Rt=0.76 min; MS m/z [M+H]⁺ 660.2, m/z [M−H]⁻ 658.3; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.23 (s, br, 1H), 10.10 (s, 1H), 10.04 (s,br, 1H), 8.55 (s, 1H), 7.78 (d, J=13.1 Hz, 1H), 7.71 (d, J=7.8 Hz, 1H),6.81 (m, 1H), 5.32 (s, 2H), 4.52 (m, 1H), 4.25 (m, 2H), 3.80 (m, 2H),3.50 (m, 3H), 3.25 (m, 1H), 2.98 (m, 3H), 2.82 (m, 1H), 2.65 (m, 1H),2.50 (m, 2H), 2.43 (s, 3H), 2.36 (s, 3H), 1.18 (t, J=7.1 Hz, 3H)

Example 55:2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

3-Hydroxypicolinic acid (170 mg, 1.20 mmol) was dissolved in DCM (6 mL)at RT under argon. 1-Chloro-N,N,2-trimethylprop-1-en-1-amine (180 mg,1.32 mmol) was added and the RM was stirred at RT for 1.5 hours. Asolution of2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide(Intermediate AD) (400 mg, 600 μmol) in DCM (4 mL) and DIPEA (524 μL,3.00 mmol) was added. The resulting brown solution was stirred at RT for2.3 hours. The RM was quenched with water (5 mL) and aq sat NaHCO₃ (5mL). It was extracted with DCM (4×40 mL). The organic layers werecombined, washed with aq sat NaHCO₃ and water, dried through a phaseseparator and concentrated under reduced pressure. The residue wasadsorbed onto Isolute and purified by column chromatography (RediSepColumn: Silica 24 g, eluent DCM:MeOH 100:0 to 90:10). The productcontaining fractions were combined and concentrated. The resulting beigesolid was purified by SFC (SFC 6) to give the title compound.

LC-MS: Rt=0.94 min; MS m/z [M+H]⁺ 668.4, m/z [M−H]⁻ 666.2; UPLC-MS 3

¹H NMR (600 MHz, DMSO-d₆) δ 10.40 (s, 1H), 10.23 (s, 1H), 8.21 (d, J=8.4Hz, 1H), 8.07 (m, 1H), 7.74 (d, J=8.4 Hz, 1H), 7.29 (m, 2H), 5.87 (m,1H), 5.28 (s, 2H), 4.55 (m, 1H), 4.11 (m, 2H), 3.48 (m, 2H), 3.40 (m,1H), 3.22 (m, 1H), 2.98 (m, 3H), 2.80 (m, 1H), 2.62 (m, 1H), 2.59 (s,3H), 2.19 (m, 2H), 1.85 (m, 2H), 1.19 (t, J=7.4 Hz, 3H)

Example 56:2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

To the stirred solution of 4-chloro-3-hydroxypicolinic acid (381 mg,2.20 mmol) in DCM (20 mL) at RT, were added PyAOP (1.15 g, 2.20 mmol),DIPEA (575 μL, 3.29 mmol) and then2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide(Intermediate AD) (600 mg, 1.10 mmol), and the RM was stirred at RT for14 hours. Water (20 mL) was added to the RM and it was extracted with10% MeOH in DCM (2×20 mL). The organic layers were combined, washed withbrine (20 mL), dried over Na₂SO₄ and concentrated. The crude product waspurified by column chromatography (silica gel column: silica 12 g,eluent DCM:MeOH 100:0 to 93:7). Then it was further purified by reversephase preparative HPLC (RP-HPLC acidic 8: 25 to 35% B in 2 min, 35 to75% B in 10 min) to give after lyophilization the title compound as awhite solid.

LC-MS: Rt=0.96 min; MS m/z [M+H]⁺ 702.3, m/z [M−H]⁻ 700.3; UPLC-MS 1

LC-MS: Rt=4.75 min; MS m/z [M+H]⁺ 702.2, m/z [M−H]⁻ 700.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.8 (m, br, 1H), 10.23 (s, 1H), 8.21 (d,J=8.4 Hz, 1H), 8.03 (d, J=4.7 Hz, 1H), 7.73 (d, J=8.4 Hz, 1H), 7.54 (d,J=4.8 Hz, 1H), 5.86 (m, 1H), 5.28 (s, 2H), 4.54 (m, 1H), 4.10 (m, 2H),3.52 (m, 3H), 3.24 (m, 1H), 2.99 (m, 3H), 2.81 (m, 1H), 2.64 (m, 1H),2.58 (s, 3H), 2.18 (m, 2H), 1.84 (m, 2H), 1.19 (t, J=7.2 Hz, 3H)

Example 57:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Step 1:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To a stirred solution of2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AP) (8.30 g, 15.2 mmol),5-methoxy-6-methylpyrimidine-4-carboxylic acid (Intermediate CW) (3.46g, 19.8 mmol) and HATU (8.68 g, 22.8 mmol) in DMF (80 mL) was addedDIPEA (13.3 mL, 76.0 mmol) at RT and the RM was stirred at RT for 15minutes. The RM was diluted with EtOAc/water, extracted twice with EtOAcand the combined organic extracts were dried over Na₂SO₄ andconcentrated. The crude product was purified by column chromatography(RediSep Column: Silica 120 g, eluent DCM:DCM/MeOH (8/2) 100:0 to65:35). The product containing fractions were combined and concentratedto give the title compound as a beige foam. Tituration of the product inEt₂O at 40° C. for 30 minutes afforded the title compound as a whitesolid.

LC-MS: Rt=1.02 min; MS m/z [M+H]⁺ 696.3, m/z [M−H]⁻ 694.5; UPLC-MS 1

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To a stirred solution of2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(8.46 g, 8.63 mmol) in DMF (80 mL) was added LiCl (1.46 g, 34.5 mmol) atRT and the RM was stirred at 210° C. for 12 minutes in the MW (splitinto 8 vials). The RM was diluted with EtOAc/water, the pH was adjustedto 2-3 with 1N HCl, and the mixture was extracted 3 times with EtOAc.The combined organic extracts were washed with brine, dried over Na₂SO₄and concentrated. The crude product was purified by columnchromatography (RediSep Column: Silica 120 g Gold, eluent DCM:DCM/MeOH(8/2) 100:0 to 70:30). The product containing fractions were combinedand concentrated. The solid was triturated in Et₂O for 30 minutes at 40°C. to give the title compound as a colorless solid characterized by theXRPD diffractogram in FIG. 4 . The table below shows the most prominentpeaks (deg 2theta) of the XRPD diffractogram of FIG. 4 .

The sodium salt was prepared analogous to the general procedure.

LC-MS: Rt=1.00 min; MS m/z [M+H]⁺ 682.5, m/z [M−H]⁻ 680.6; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.34 (s, br, 1H), 10.00 (s, 1H), 8.55 (s,1H), 7.72 (d, J=8.5 Hz, 1H), 7.62 (m, 1H), 7.53 (d, J=8.8 Hz, 1H), 6.83(m, 1H), 5.24 (s, 2H), 4.52 (m, 1H), 4.26 (m, 2H), 3.81 (m, 2H), 3.50(m, 3H), 3.25 (m, 1H), 3.00 (m, 3H), 2.81 (m, 1H), 2.63 (m, 1H), 2.51(m, 2H), 2.43 (s, 3H), 2.35 (s, 3H), 1.20 (t, J=7.4 Hz, 3H)

Angle d Value 2-Theta° Angstrom Intensity 11.81 7.49 medium 13.75 6.44medium 14.45 6.13 medium 15.32 5.78 medium 17.04 5.20 high 17.40 5.09medium 18.27 4.85 medium 19.95 4.45 medium 22.92 3.88 medium 27.13 3.28medium

Example 58:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

3-Hydroxypicolinic acid (110 mg, 778 μmol) was dissolved in DCM (5 mL)at RT under argon. 1-Chloro-N,N,2-trimethylprop-1-en-1-amine (113 μL,856 μmol) was added and the RM was stirred at RT for 1.75 hours. Asolution of2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AP) (312 mg, 389 μmol) in DCM (2 mL) and DIPEA (340 μL,1.94 mmol) were added and the RM was stirred at RT for 2.5 hours. The RMwas quenched with water (5 mL) and aq sat NaHCO₃ (5 mL) and extractedwith DCM (4×20 mL). The combined organic layers were washed with aq satNaHCO₃ (5 mL) and water, dried over a phase separator and concentratedunder reduced pressure. The crude product was adsorbed onto Isolute andpurified by column chromatography (RediSep Column: Silica 24 g, eluentDCM:MeOH 100:0 to 90:10). The product containing fractions were combinedand concentrated under reduced pressure. The still impure product waspurified in 2 portions by reverse phase preparative HPLC (RP-HPLC acidic1: 10 to 90% B in 20 min and RP-HPLC acidic 1: 20 to 80% B in 20 min).The product containing fractions were combined, basified with aq satNaHCO₃ and the ACN was removed under reduced pressure. The residue wasextracted with DCM (3×10 mL), dried through a phase separator andconcentrated under reduced pressure to give the title compound as a paleyellow solid.

The sodium salt was prepared analogous to the general procedure.

LC-MS: Rt=1.00 min; MS m/z [M+H]⁺ 667.4, m/z [M−H]⁻ 665.4; UPLC-MS 4

¹H NMR (400 MHz, DMSO-d₆) δ 10.39 (s, 1H), 10.01 (s, 1H), 8.06 (m, 1H),7.72 (d, J=8.4 Hz, 1H), 7.63 (m, 1H), 7.53 (d, J=8.6 Hz, 1H), 7.29 (m,2H), 6.83 (m, 1H), 5.24 (s, 2H), 4.55 (m, 1H), 4.26 (m, 2H), 3.81 (m,2H), 3.44 (m, 3H), 3.22 (m, 1H), 2.99 (m, 3H), 2.80 (m, 1H), 2.62 (m,1H), 2.54 (m, 2H), 2.35 (s, 3H), 1.20 (t, J=7.4 Hz, 3H)

Example 59:2-(2-(3,4-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

2-(2-(3,6-Dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Example 58) (100 mg, 145 μmol) was suspended in DMF (500 μL). Sodiummethanethiolate (100 mg, 1.43 mmol) was added and the RM was stirred at120° C. for 40 minutes. The RM was allowed to cool to RT and it waspartitioned between water (60 mL) and DCM/MeOH (3:1) (50 mL). Theorganic layer was separated, dried over MgSO₄, filtered and reduced invacuo to give a solution of the crude product in DMF, which was purifiedin 2 portions by reverse phase preparative HPLC (RP-HPLC acidic 1: 30 to55% B in 15 min with a plateau at 55% for 1 min and RP-HPLC acidic 1: 30to 50% B in 15 min with a plateau at 50% for 1 min). The productcontaining fractions were basified with 5% aq NaHCO₃ and extracted withDCM (2×10 mL). The organics were eluted through a phase separator andevaporated in vacuo to give the title compound as a white solid.

LC-MS: Rt=1.02 min; MS m/z [M+H]⁺ 667.5, m/z [M−H]⁻ 665.5; UPLC-MS 1

LC-MS: Rt=3.02 min; MS m/z [M+H]⁺ 667.4, m/z [M−H]⁻ 665.3; UPLC-MS 9

¹H NMR (600 MHz, DMSO-d₆) δ 10.38 (s, br, 1H), 9.99 (s, 1H), 8.06 (m,1H), 7.68 (d, J=8.2 Hz, 1H), 7.62 (m, 1H), 7.53 (d, J=8.2 Hz, 1H), 7.28(m, 2H), 6.51 (m, 1H), 5.22 (s, 2H), 4.83 (m, 1H), 4.54 (m, 1H), 4.08(m, 1H), 4.01 (m, 1H), 3.62 (m, 1H), 3.46 (m, 3H), 3.22 (m, 1H), 2.95(m, 3H), 2.78 (m, 1H), 2.59 (m, 1H), 2.34 (s, 3H), 2.09 (m, 2H), 1.18(t, J=7.5 Hz, 3H)

Example 60:2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-2-(6-oxo-3,6-dihydro-2H-pyran-4-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To a 3 L plastic Erlenmeyer covered with a breathable seal was addedMcllvaine Puffer pH 4.5 (970 mL),2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Example 58) (100 mg, 150 μmol) dissolved in DMSO (5 mL),N-hydroxyphthalimide (163 mg, 1.00 mmol) dissolved in DMSO (5 mL) andlaccase (3.00 g) (Supplier: Aldrich laccase from trametes versicolor 0.5U/mg). The RM was stirred at 30° C. 150 RPM in an Infors HT MultitronShaker. After 3 hours, the conversion stopped at 19% and laccase (1.00g) was added. A decrease in the conversion was observed in the next 45minutes and the reaction was quenched and extracted with EtOAc (1 L) andthen EtOAc (500 mL). The organic layer was dried over MgSO₄ andconcentrated to dryness. The crude product was dissolved in DMSO (4 mL)and purified by reverse phase preparative HPLC (RP-HPLC acidic 13: 0 to70% B in 30 min with a plateau at 70% for 5 min). The target compoundwas dried and submitted to SFC (SFC 7) to give the title compound.

LC-MS: Rt=0.94 min; MS m/z [M+H]⁺ 681.4, m/z [M−H]⁻ 679.4; UPLC-MS 1

¹H NMR (600 MHz, DMSO-d₆) δ 10.08 (s, 1H), 8.05 (m, 1H), 7.70 (d, J=8.4Hz, 1H), 7.63 (m, 1H), 7.53 (d, J=8.4 Hz, 1H), 7.28 (m, 2H), 6.66 (m,1H), 5.29 (s, 2H), 4.52 (m, 3H), 3.46 (m, 3H), 3.23 (m, 1H), 3.03 (m,2H), 2.97 (m, 3H), 2.81 (m, 1H), 2.63 (m, 1H), 2.36 (s, 3H), 1.20 (t,J=7.5 Hz, 3H)

Example 61:2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

4-Chloro-3-hydroxypicolinic acid (199 mg, 1.15 mmol) was dissolved inDCM (12 mL) at RT under argon. 1-Chloro-N,N,2-trimethylprop-1-en-1-amine(171 mg, 1.28 mmol) was added and the RM was stirred at RT for 2.5hours.2-(2-(3,6-Dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AP) (250 mg, 458 μmol) and DIPEA (320 μL, 1.83 mmol) wereadded and the brown solution was stirred at RT for 1.25 hours. The RMwas quenched with water (10 mL) and aq sat NaHCO₃ (10 mL). Then it wasextracted with DCM (4×40 mL). The organic layers were combined andwashed with water (10 mL), dried through a phase separator andconcentrated under reduced pressure. The residue was adsorbed ontoIsolute and purified by column chromatography (RediSep Column: Silica 24g, eluent DCM:MeOH 100:0 to 80:20). The product containing fractionswere combined, concentrated under reduced pressure and dried under HV togive the title compound as a beige solid. A part of the solid wasdissolved in MeOH (2.5 mL) and DCM (2.5 mL) and crystallized onstanding. The resulting solid was dried under HV to give the titlecompound.

LC-MS: Rt=1.09 min; MS m/z [M+H]⁺ 701.5, m/z [M−H]⁻ 699.3; UPLC-MS 1

¹H NMR (600 MHz, DMSO-d₆) δ 10.83 (s, 1H), 10.00 (s, 1H), 8.06 (d, J=5.0Hz, 1H), 7.72 (d, J=8.4 Hz, 1H), 7.62 (m, 1H), 7.56 (d, J=5.1 Hz, 1H),7.53 (d, J=8.2 Hz, 1H), 6.83 (m, 1H), 5.24 (s, 2H), 4.54 (m, 1H), 4.26(m, 2H), 3.81 (m, 2H), 3.53 (m, 3H), 3.25 (m, 1H), 3.00 (m, 3H), 2.81(m, 1H), 2.64 (m, 1H), 2.54 (m, 2H), 2.35 (s, 3H), 1.20 (t, J=7.5 Hz,3H)

Example 62:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-fluoro-3-hydroxy-6-methylpicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Step 1:2-(6-(4-(3-(benzyloxy)-4-fluoro-6-methylpicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

2-(2-(3,6-Dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AP) (285 mg, 522 μmol),3-(benzyloxy)-4-fluoro-6-methylpicolinic acid (Intermediate CX) (171 mg,575 μmol) and HATU (218 mg, 575 μmol) were suspended in DCM (5 mL) andcooled to 0° C. Then DIPEA (228 μL, 1.31 mmol) was added and the RM wasstirred at RT for 2.5 hours. Water (20 mL), aq sat NaHCO₃ (10 mL) andDCM (30 mL) were added. The aqueous layer was washed with DCM (2×20 mL).The combined organic layers were dried through a phase separator andconcentrated under reduced pressure. The crude product was purified in 2portions by reverse phase preparative HPLC (2×RP-HPLC basic 1: 5 to 95%B in 20 min with a plateau at 95% for 1 min). The product containingfractions were combined, basified with aq sat NaHCO₃, extracted twicewith DCM, dried through a phase separator and concentrated under reducedpressure to give the title compound.

LC-MS: Rt=1.23 min; MS m/z [M+H]⁺ 789.6, m/z [M−H]⁻ 787.3; UPLC-MS 1

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-fluoro-3-hydroxy-6-methylpicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

2-(6-(4-(3-(Benzyloxy)-4-fluoro-6-methylpicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(156 mg, 178 μmol) was dissolved in DCM (5 mL) and TFA (5.00 mL, 64.9mmol) was added and the RM was stirred at 60° C. overnight. The RM wasconcentrated under reduced pressure. The crude product was purified byreverse phase preparative ISCO (RediSep Column: C18 50 g Gold, eluentwater+0.1% TFA:ACN 100:0 to 0:100). The product containing fractionswere combined, basified with aq sat NaHCO₃, extracted twice with DCM,dried through a phase separator and concentrated under reduced pressureto give the title compound.

LC-MS: Rt=1.02 min; MS m/z [M+H]⁺ 699.5, m/z [M−H]⁻ 697.5; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.36 (s, 1H), 9.99 (s, 1H), 7.72 (d, J=8.4Hz, 1H), 7.62 (m, 1H), 7.53 (d, J=8.6 Hz, 1H), 7.22 (d, J=11.6 Hz, 1H),6.83 (m, 1H), 5.24 (s, 2H), 4.53 (m, 1H), 4.26 (m, 2H), 3.81 (m, 2H),3.47 (m, 3H), 3.23 (m, 1H), 3.00 (m, 3H), 2.80 (m, 1H), 2.63 (m, 1H),2.52 (m, 2H), 2.39 (s, 3H), 2.35 (s, 3H), 1.20 (t, J=7.4 Hz, 3H)

Example 63:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-fluoro-3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Step 1:2-(6-(4-(3-(benzyloxy)-4-fluoropicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To a stirred solution of2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AP) (500 mg, 916 μmol), 3-(benzyloxy)-4-fluoropicolinicacid (Intermediate CU) (238 mg, 962 μmol) and HATU (418 mg, 1.10 mmol)in DMF (10 mL) was added DIPEA (800 μL, 4.58 mmol) at RT and the RM wasstirred at RT for 10 minutes. The RM was diluted with EtOAc/water,extracted twice with EtOAc and the combined organic extracts were driedover Na₂SO₄ and concentrated. The crude product was purified by columnchromatography (RediSep Column: Silica 24 g, eluent DCM:MeOH 100:0 to40:60). The product containing fractions were combined and concentratedto give the title compound as a white foam.

LC-MS: Rt=1.20 min; MS m/z [M+H]⁺ 775.3, m/z [M−H]⁻ 773.5; UPLC-MS 1

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-fluoro-3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To a stirred suspension of2-(6-(4-(3-(benzyloxy)-4-fluoropicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(510 mg, 652 μmol) in DCM (10 mL) was added boron trichloride methylsulfide complex (652 μL, 1.30 mmol) at RT and the RM was stirred at RTfor 14 hours. The RM was quenched with MeOH. Then it was diluted withDCM/water, extracted twice with DCM and the combined organic extractswere washed with water and brine, dried over Na₂SO₄ and concentrated.The crude product was purified by column chromatography (RediSep Column:Silica 24 g, eluent DCM:MeOH 100:0 to 20:80). The product containingfractions were combined and concentrated. The resulting solid waspurified by reverse phase preparative HPLC (RP-HPLC acidic 1: 15 to 75%B in 20 min with a plateau at 75% for 1 min). The combined fractionswere basified with NaHCO₃, the ACN was evaporated and the resultingaqueous layer was extracted with DCM, the organic extract was washedwith brine, dried over Na₂SO₄ and concentrated to give the titlecompound as a white solid.

The sodium salt was prepared analogous to the general procedure.

LC-MS: Rt=0.98 min; MS m/z [M+H]⁺ 685.3, m/z [M−H]⁻ 683.3; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.76 (s, br, 1H), 10.00 (s, 1H), 8.06 (m,1H), 7.72 (d, J=8.4 Hz, 1H), 7.62 (m, 1H), 7.53 (d, J=8.6 Hz, 1H), 7.34(m, 1H), 6.83 (m, 1H), 5.24 (s, 2H), 4.54 (m, 1H), 4.26 (m, 2H), 3.81(m, 2H), 3.46 (m, 3H), 3.23 (m, 1H), 3.00 (m, 3H), 2.81 (m, 1H), 2.62(m, 1H), 2.50 (m, 2H), 2.35 (s, 3H), 1.19 (t, J=7.4 Hz, 3H)

Example 64:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxypyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Step 1:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxypyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To a beige suspension of 5-methoxy-pyrimidine-4-carboxylic acid (83.0mg, 522 μmol) in DCM (6 mL) was added1-chloro-N,N,2-trimethyl-1-prop-1-en-1-amine (120 μL, 871 μmol). The RMwas stirred at RT for 5 minutes. DIPEA (182 μL, 1.05 mmol) was addedslowly, followed by2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AP) (200 mg, 348 μmol). The RM became progressively ayellow to brown solution and was stirred at RT for 28 hours. There wasstill starting material left. To a beige suspension of5-methoxy-pyrimidine-4-carboxylic acid (83.0 mg, 522 μmol) in DCM (6 mL)was added 1-chloro-N,N,2-trimethyl-1-prop-1-en-1-amine (120 μL, 871μmol). The RM was stirred at RT for 5 minutes. DIPEA (182 μL, 1.05 mmol)was added, followed by the previous RM. The mixture was stirred at RTfor 18 hours. There was still some starting material left, as detectedby LCMS. To a beige suspension of 5-methoxy-pyrimidine-4-carboxylic acid(83.0 mg, 522 μmol) in DCM (6 mL) was added1-chloro-N,N,2-trimethyl-1-prop-1-en-1-amine (120 μL, 871 μmol). The RMwas stirred at RT for 30 minutes. DIPEA (182 μL, 1.05 mmol) was addedslowly, followed by the previous RM. The resulting beige/light yellow RMwas stirred at RT for 4 hours, diluted with DCM and washed with water.The aqueous layer was extracted twice with DCM. The combined organiclayers were dried through a phase separator, concentrated and driedunder reduced pressure to give a brown residue. The crude product waspurified by column chromatography (RediSep Column: Silica 12 g, eluentDCM:DCM/MeOH (9/1) 100:0 to 50:50). The product containing fractionswere combined, concentrated and dried under reduced pressure to give thetitle compound as a beige solid.

LC-MS: Rt=0.93 min; MS m/z [M+H]⁺ 682.4, m/z [M−H]⁻ 680.5; UPLC-MS 1

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxypyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To a light yellow solution of2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxypyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(146 mg, 176 μmol) in DMF (1.8 mL) was added LiCl (22.3 mg, 527 μmol).The light yellow RM was heated to 145° C. and stirred for 2 hours. LiCl(22.3 mg, 527 μmol) was added, and the brown RM was heated to 145° C.and stirred for 21 hours. The dark brown RM was diluted with DCM andwashed once with water. The aqueous layer was extracted four times withDCM and a small amount of MeOH. The combined organic layers were driedthrough a phase separator and concentrated to give the crude product asa solution in remaining DMF, which was purified by reverse phasepreparative HPLC (RP-HPLC basic 1: 0 to 20% B in 27 min with a plateauat 20% for 1 min). The product containing fractions were combined,concentrated and lyophilized to give the title compound.

LC-MS: Rt=0.93 min; MS m/z [M+H]⁺ 668.4, m/z [M−H]⁻ 666.5; UPLC-MS 1

¹H NMR (600 MHz, DMSO-d₆) δ 10.01 (s, 1H), 8.60 (s, 1H), 8.37 (s, 1H),7.71 (d, J=8.5 Hz, 1H), 7.62 (m, 1H), 7.53 (d, J=8.4 Hz, 1H), 6.83 (m,1H), 5.24 (s, 2H), 4.51 (m, 1H), 4.26 (m, 2H), 3.80 (m, 2H), 3.47 (m,3H), 3.25 (m, 1H), 2.99 (m, 3H), 2.80 (m, 1H), 2.62 (m, 1H), 2.50 (m,2H), 2.35 (s, 3H), 1.19 (t, J=7.5 Hz, 3H)

Example 65:2-(6-(4-(5-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

5-Chloro-3-hydroxypicolinic acid (300 mg, 1.73 mmol) was dissolved inDCM (5 mL) and 1-chloro-N,N,2-trimethylprop-1-en-1-amine (301 μL, 2.27mmol) was added. The mixture was stirred at RT for 2 hours. Then2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AP) (100 mg, 152 μmol) was added. The mixture was stirredat RT for 1 hour. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL)were added. The aqueous layer was washed with DCM (2×10 mL). Thecombined organic layers were dried through a phase separator andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (RediSep Column: Silica 40 g, eluent DCM:DCM/MeOH(9/1) 100:0 to 80:20). The product containing fractions were combined,concentrated and dried under reduced pressure to give impure productwhich was further purified by reverse phase preparative HPLC (RP-HPLCacidic 1: 5 to 100% B in 20 min). The product containing fractions werecombined, basified with aq sat NaHCO₃, extracted twice with DCM, driedthrough a phase separator and concentrated under reduced pressure togive the title compound.

LC-MS: Rt=1.08 min; MS m/z [M+H]⁺ 701.4, m/z [M−H]⁻ 699.4; UPLC-MS 3

¹H NMR (400 MHz, DMSO-d₆) δ 10.97 (s, 1H), 10.00 (s, 1H), 8.13 (d, J=2.0Hz, 1H), 7.72 (d, J=8.4 Hz, 1H), 7.63 (m, 1H), 7.53 (d, J=8.34 Hz, 1H),7.37 (d, J=2.0 Hz, 1H), 6.83 (m, 1H), 5.24 (s, 2H), 4.53 (m, 1H), 4.27(m, 2H), 3.81 (m, 2H), 3.44 (m, 3H), 3.22 (m, 1H), 2.99 (m, 3H), 2.80(m, 1H), 2.61 (m, 1H), 2.51 (m, 2H), 2.35 (s, 3H), 1.20 (t, J=7.5 Hz,3H)

Example 66:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxy-4-methylpicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To a suspension of 3-hydroxy-4-methylpyridine-2-carboxylic acid.HCl(56.1 mg, 281 μmol) in DCM (3.1 mL) was added1-chloro-N,N,2-trimethylprop-1-en-1-amine (60.0 μL, 435 μmol). The RMwas stirred at RT for 55 minutes, and DIPEA (152 μL, 871 μmol) was addedslowly, followed by2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AP) (100 mg, 174 μmol). The RM was stirred at RT for 18.25hours, diluted with DCM and washed with water. The aqueous layer wasextracted twice with DCM. The combined organic layers were dried througha phase separator and concentrated under reduced pressure. The crudeproduct was suspended in MeOH and filtered. The filtrate wasconcentrated under reduced pressure, and the residue was purified byreverse phase preparative HPLC (RP-HPLC acidic 1: 20 to 50% B in 20 min,with a plateau at 50% for 1 min). The product containing fractions werecombined and concentrated to remove the ACN. The residue was basifiedwith solid NaHCO₃, extracted twice with DCM, dried through a phaseseparator and concentrated under reduced pressure to give the titlecompound.

LC-MS: Rt=1.05 min; MS m/z [M+H]⁺ 681.5, m/z [M−H]⁻ 679.4; UPLC-MS 3

¹H NMR (600 MHz, DMSO-d₆) δ 10.39 (s, br, 1H), 10.02 (s, 1H), 7.98 (d,J=4.6 Hz, 1H), 7.72 (d, J=8.4 Hz, 1H), 7.62 (m, 1H), 7.53 (d, J=8.4 Hz,1H), 7.25 (d, J=4.6 Hz, 1H), 6.83 (m, 1H), 5.25 (s, 2H), 4.57 (m, 1H),4.26 (m, 2H), 4.10 (m, 1H), 3.81 (m, 2H), 3.51 (m, 2H), 3.29 (m 1H),3.01 (m, 3H), 2.82 (m, 1H), 2.66 (m, 1H), 2.52 (m, 2H), 2.35 (s, 3H),2.23 (s, 3H), 1.20 (t, J=7.5 Hz, 3H)

Example 67:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxy-4-(trifluoromethoxy)picolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Step 1:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-methoxy-4-(trifluoromethoxy)picolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To a suspension of 3-methoxy-4-(trifluoromethoxy)picolinic acid(Intermediate CZ) (32.9 mg, 136 μmol) in DCM (1.9 mL) was added1-chloro-N,N,2-trimethylprop-1-en-1-amine (39.0 μL, 283 μmol), and theRM was stirred at RT for 1 hour. DIPEA (59.3 μL, 340 μmol) was addedslowly, followed by2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AP) (65.0 mg, 113 μmol). The RM was stirred at RT for 3hours, diluted with DCM and washed with water. The aqueous layer wasextracted twice with DCM. The combined organic layers were dried througha phase separator and concentrated under reduced pressure. The crudeproduct was purified by column chromatography (RediSep Column: Silica 4g, eluent DCM:DCM/MeOH (8/2) 100:0 to 50:50). The product containingfractions were combined and concentrated under reduced pressure to givethe title compound as a beige solid.

LC-MS: Rt=1.17 min; MS m/z [M+H]⁺ 765.5, m/z [M−H]⁻ 763.5; UPLC-MS 1

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxy-4-(trifluoromethoxy)picolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

A RM of2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-methoxy-4-(trifluoromethoxy)picolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(48.0 mg, 49.0 μmol) and LiCl (8.30 mg, 196 μmol) in DMF (490 μL) wassubmitted to MW irradiations (very high absorption) at 200° C. for 10minutes. The RM was diluted with water and extracted three times withEtOAc. The combined organic layers were washed once with brine, driedover Na₂SO₄, concentrated and dried under HV. The residue was purifiedby reverse phase preparative HPLC (RP-HPLC basic 1: 5 to 70% B in 20min, with a plateau at 70% for 1 min). The product containing fractionswere combined and concentrated to remove the ACN, then it waslyophilized to give the title compound.

LC-MS: Rt=1.17 min; MS m/z [M+H]⁺ 751.3, m/z [M−H]⁻ 749.3; UPLC-MS 1

Example 68:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-2-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Step 1:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxy-2-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To a brown suspension of 5-methoxy-2-methylpyrimidine-4-carboxylic acid(40.6 mg, 229 μmol) in DCM (3.1 mL) was added1-chloro-N,N,2-trimethylprop-1-en-1-amine (35.4 μL, 257 μmol). The RMwas stirred at RT for 30 minutes. DIPEA (96.0 μL, 550 μmol) was addedslowly, followed by2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AP) (100 mg, 174 μmol). The brown solution was stirred atRT for 20 hours. To a suspension of5-methoxy-2-methylpyrimidine-4-carboxylic acid (40.6 mg, 229 μmol) inDCM (3.1 mL) was added 1-chloro-N,N,2-trimethylprop-1-en-1-amine (35.4μL, 257 μmol). The RM was stirred at RT for 2.25 hours. DIPEA (96.0 μL,550 μmol) was added slowly, followed by the previous RM. The RM wasstirred at RT over the weekend. The RM was diluted with DCM and washedwith water. The aqueous layer was extracted twice with DCM. The combinedorganic layers were dried through a phase separator and concentratedunder reduced pressure. The aqueous layer still contained product so itwas extracted twice with DCM and a small amount of MeOH. The combinedorganic layers were dried through a phase separator and concentratedunder reduced pressure. The first crude product was purified by columnchromatography (RediSep Column: Silica 4 g, eluent EtOAc:EtOAc/EtOH(95/5) 100:0 to 0:100). The product containing fractions were combinedand concentrated to give product. The second crude product was purifiedby column chromatography (RediSep Column: Silica 4 g, eluentEtOAc:EtOAc/EtOH (95/5) 100:0 to 0:100). The product containingfractions were combined and concentrated to give product. Both materialswere combined to give the title compound.

LC-MS: Rt=0.97 min; MS m/z [M+H]⁺ 696.5, m/z [M−H]⁻ 694.4; UPLC-MS 3

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-2-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To a light yellow solution of2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxy-2-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(28.0 mg, 38.0 μmol) in DMF (396 μL) was added LiCl (5.04 mg, 119 μmol).The RM was heated to 145° C. and stirred for 7.5 hours. LiCl (3.00 mg,71.0 μmol) was added. The RM was heated to 145° C. and stirred for 15hours. The RM was diluted with DCM and washed once with water. Theaqueous layer was extracted once with DCM. The combined organic layerswere dried through a phase separator, concentrated and dried undervacuum. The residue was purified by reverse phase preparative HPLC(RP-HPLC basic 1: 5 to 50% B in 20 min, with a plateau at 50% for 1min). The product containing fraction was lyophilized to give the titlecompound.

LC-MS: Rt=0.58 min; MS m/z [M+H]⁺ 682.5, m/z [M−H]⁻ 680.4; UPLC-MS 5

¹H NMR (600 MHz, DMSO-d₆) δ 10.41 (s, br, 1H), 10.01 (s, 1H), 8.33 (s,1H), 7.72 (d, J=8.4 Hz, 1H), 7.63 (m, 1H), 7.54 (d, J=8.1 Hz, 1H), 6.83(m, 1H), 5.25 (s, 2H), 4.51 (m, 1H), 4.27 (m, 2H), 3.81 (m, 2H), 3.47(m, 3H), 3.25 (m, 1H), 2.99 (m, 3H), 2.81 (m, 1H), 2.62 (m, 1H), 2.53(s, 3H), 2.50 (m, 2H), 2.36 (s, 3H), 1.20 (t, J=7.5 Hz, 3H)

Example 69:N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AQ) (170 mg, 293 μmol) was suspended in DMF (5 mL) and3-hydroxypicolinic acid (82.0 mg, 586 μmol), DIPEA (256 μL, 1.47 mmol),HOBt (79.0 mg, 586 μmol) and EDC.HCl (112 mg, 586 μmol) were added tothe RM and it was stirred at RT for 12 hours. Water was added to the RMand it was filtered to obtain a solid crude product which was purifiedby reverse phase preparative HPLC (RP-HPLC basic 2: 15 to 25% B in 2min, 25 to 60% B in 10 min) to give the title compound.

LC-MS: Rt=1.05 min; MS m/z [M+H]⁺ 701.6, m/z [M−H]⁻ 699.4; UPLC-MS 1

LC-MS: Rt=5.22 min; MS m/z [M+H]⁺ 701.2, m/z [M−H]⁻ 699.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.40 (s, br, 1H), 10.11 (s, br, 1H), 8.06(m, 1H), 7.96 (s, 1H), 7.75 (s, 1H), 7.28 (m, 2H), 6.82 (m, 1H), 5.27(s, 2H), 4.55 (m, 1H), 4.25 (m, 2H), 3.81 (m, 2H), 3.44 (m, 3H), 3.22(m, 1H), 2.98 (m, 3H), 2.80 (m, 1H), 2.62 (m, 1H), 2.50 (m, 2H), 2.35(s, 3H), 1.18 (t, J=6.8 Hz, 3H)

Example 70:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

2-(2-(3,6-Dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide(Intermediate AR) (1.40 g, 1.84 mmol) was dissolved in DCM (10 mL) and3-hydroxypicolinoyl chloride (Intermediate CV) (520 mg, 3.30 mmol) wasadded, followed by DIPEA (1.60 mL, 9.18 mmol). The RM was stirred at RTfor 2 hours. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL) wereadded. The aqueous layer was washed with DCM (2×10 mL). The combinedorganic layers were dried through a phase separator and concentratedunder reduced pressure. The crude product was purified by columnchromatography (RediSep Column: Silica 40 g, eluent DCM:DCM/MeOH (9/1)100:0 to 30:70). The product containing fractions were combined,concentrated under vacuum and dried under HV to afford a grey solidwhich was purified by reverse phase preparative HPLC (RP-HPLC acidic 1:5 to 95% B in 20 min). The product containing fractions were combined,basified with aq sat NaHCO₃, extracted twice with DCM, dried through aphase separator and concentrated under reduced pressure to give thetitle compound.

LC-MS: Rt=0.96 min; MS m/z [M+H]⁺ 686.4, m/z [M−H]⁻ 684.4; UPLC-MS 3

¹H NMR (400 MHz, DMSO-d₆) δ 10.38 (s, 1H), 10.33 (s, br, 1H), 8.31 (d,J=12.4 Hz, 1H), 8.07 (m, 1H), 7.29 (m, 2H), 6.82 (m, 1H), 5.33 (s, 2H),4.55 (m, 1H), 4.26 (m, 2H), 3.81 (m, 2H), 3.46 (m, 3H), 3.23 (m, 1H),2.97 (m, 3H), 2.80 (m, 1H), 2.62 (m, 1H), 2.59 (s, 3H), 2.51 (m, 2H),1.19 (t, J=7.5 Hz, 3H)

Example 71:(R)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)-3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

3-Hydroxypicolinic acid (59.8 mg, 421 μmol) was dissolved in DCM (3.4mL) at RT under argon. 1-Chloro-N,N,2-trimethylprop-1-en-1-amine (61.3μL, 463 μmol) was added and the RM was stirred at RT for 1.5 hours. Asolution of(R)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide(Intermediate BF) (174 mg, 210 μmol) in DCM (1 mL) and DIPEA (184 μL,1.05 mmol) was added and the brown solution was stirred at RT for 24hours. The RM was quenched with water (5 mL) and aq sat NaHCO₃ (5 mL)and then extracted with DCM (4×20 mL). The organic layers were combined,dried through a phase separator and concentrated under reduced pressure.The residue was adsorbed onto Isolute and purified by columnchromatography (RediSep Column: Silica 40 g, eluent DCM:MeOH 100:0 to90:10). The product containing fractions were combined and concentratedunder reduced pressure. This material was purified by reverse phasepreparative HPLC (RP-HPLC acidic 1: 10 to 90% B in 20 min). The productcontaining fractions were lyophilized. The resulting solid was dissolvedin DCM/MeOH and filtered through a PL-HCO₃ MP SPE (100 mg per 6 mL)cartridge. The filtrate was concentrated and dried under HV to give thetitle compound as a colorless solid.

LC-MS: Rt=1.04 min; MS m/z [M+H]⁺ 667.3, m/z [M−H]⁻ 665.1; UPLC-MS 4

¹H NMR (400 MHz, DMSO-d₆) δ 10.87 (s, 1H), 10.34 (d, J=9.3 Hz, 1H), 8.07(m, 1H), 7.79 (d, J=8.5 Hz, 2H), 7.71 (d, J=8.5 Hz, 2H), 7.29 (m, 2H),6.80 (m, 1H), 5.21 (m, 2H), 4.81-4.40 (m, 1H), 4.24 (m, 2H), 3.80 (m,2H), 3.68 (m, 2H), 3.43 (m, 1H), 3.16 (m, 2H), 2.92 (m, 1H), 2.81 (m,1H), 2.63 (m, 1H), 2.55 (s, 2H), 1.39 (dd, J=6.5 Hz, 19.0 Hz, 3H), 1.20(t, J=7.2 Hz, 3H)

Example 72:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

3-Hydroxypicolinic acid (141 mg, 1.01 mmol) was dissolved in DCM (2 mL)and 1-chloro-N,N,2-trimethylprop-1-en-1-amine (221 μL, 1.67 mmol) wasadded. The RM was stirred at RT for 2 hours. Then2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide(Intermediate AS) (267 mg, 477 μmol) was added, followed by DIPEA (417μL, 2.39 mmol). The RM was stirred at RT for 2 hours. Water, aq satNaHCO₃ and DCM were added. The aqueous layer was extracted twice withDCM. The combined organic layers were dried through a phase separatorand concentrated under reduced pressure. The crude product was purifiedby reverse phase preparative HPLC (RP-HPLC acidic 1: 20 to 65% B in 30min). The product containing fractions were combined, basified with aqsat NaHCO₃, extracted twice with DCM, dried through a phase separatorand concentrated under reduced pressure. The concentrated fractions weresuspended in MeOH and sonicated. Then this material was filtered and thecake was washed three times with MeOH. The cake was purified by reversephase preparative HPLC (RP-HPLC acidic 1: 27 to 45% B in 25 min). Theproduct containing fractions were combined, basified with aq sat NaHCO₃,extracted twice with DCM, dried through a phase separator andconcentrated under reduced pressure to give the title compound.

LC-MS: Rt=0.94 min; MS m/z [M+H]⁺ 653.2, m/z [M−H]⁻ 651.1; UPLC-MS 4

¹H NMR (400 MHz, DMSO-d₆) δ 10.85 (s, 1H), 10.38 (s, 1H), 8.07 (m, 1H),7.78 (d, J=8.6 Hz, 2H), 7.70 (d, J=8.8 Hz, 2H), 7.29 (m, 2H), 6.80 (m,1H), 5.20 (s, 2H), 4.56 (m, 1H), 4.24 (m, 2H), 3.80 (m, 2H), 3.47 (m,3H), 3.23 (m, 1H), 2.98 (m, 3H), 2.81 (m, 1H), 2.63 (m, 1H), 2.51 (m,2H), 1.18 (t, J=7.4 Hz, 3H)

Example 73:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

Step 1:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

To a solution of 5-methoxy-6-methylpyrimidine-4-carboxylic acid(Intermediate CW) (74.1 mg, 423 μmol) in DMF (3 mL), was added HATU (161mg, 423 μmol). The RM was stirred for 15 minutes and then2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide(Intermediate AS) (150 mg, 282 μmol) was added, followed by DIPEA (148μL, 847 μmol).

The resulting RM was stirred at 30° C. for 16 hours. The RM was dilutedwith EtOAc and washed with water. The aqueous layer was extracted twicewith EtOAc and twice with DCM. The organic layers were combined, driedthrough a phase separator and concentrated. The crude product waspurified by column chromatography (RediSep Column: Silica 12 g, eluentDCM:MeOH 100:0 to 90:10). The product containing fractions were combinedand concentrated under reduced pressure to give the title compound as awhite solid.

LC-MS: Rt=1.02 min; MS m/z [M+H]⁺ 682.3, m/z [M−H]⁻ 680.2; UPLC-MS 3

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

A solution of2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide(70.0 mg, 103 μmol) in DMF (1 mL) under argon was cooled to −78° C. LiCl(43.5 mg, 1.03 mmol) was added portionwise. The RM was stirred at 125°C. for 26 hours. The RM was diluted with EtOAc and washed with water.The organic layer was dried through a phase separator and concentrated.The crude product was purified by reverse phase preparative HPLC(RP-HPLC basic 1: 5 to 50% B in 20 min with a plateau at 50% for 1 min).The product containing fractions were lyophilized, and the titlecompound was obtained as a colorless powder.

LC-MS: Rt=1.03 min; MS m/z [M+H]⁺ 668.5, m/z [M−H]⁻ 666.4; UPLC-MS 6

¹H NMR (400 MHz, DMSO-d₆) δ 10.87 (s, 1H), 8.55 (s, 1H), 7.78 (d, J=8.6Hz, 2H), 7.70 (d, J=8.8 Hz, 2H), 6.79 (m, 1H), 5.20 (s, 2H), 4.52 (m,1H), 4.23 (m, 2H), 3.79 (m, 2H), 3.50 (m, 3H), 3.26 (m, 1H), 2.98 (m,3H), 2.82 (m, 1H), 2.65 (m, 1H), 2.51 (m, 2H), 2.43 (s, 3H), 1.17 (t,J=7.4 Hz, 3H)

Example 74:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxy-6-methylpicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

3-Hydroxy-6-methylpicolinic acid (88.0 mg, 376 μmol) was suspended inDCM (4 mL) at RT. After cooling to 0° C.,1-chloro-N,N,2-trimethylprop-1-en-1-amine (62.0 μL, 470 μmol) was addeddropwise. The RM was stirred at RT for 2 hours, then it was cooled to 0°C. again. DIPEA (164 μL, 941 μmol) was added. This solution was addeddropwise at 0° C. to a solution of2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide(Intermediate AS) (100 mg, 188 μmol) in DCM (4 mL). The RM was stirredat RT for 18 hours. The RM was diluted with DCM, washed with aq satNH₄Cl, dried over Na₂SO₄, concentrated and dried. The crude product waspurified by reverse phase preparative HPLC (RP-HPLC basic 1: 5 to 100% Bin 20 min). The product containing fractions were combined, the ACN wasremoved and the residue was lyophilized to give the title compound as afoam.

LC-MS: Rt=1.05 min; MS m/z [M+H]⁺ 667.2, m/z [M−H]⁻ 665.1; UPLC-MS 4

¹H NMR (400 MHz, DMSO-d₆) δ 10.88 (s, 1H), 10.09 (s, br, 1H), 7.78 (d,J=8.6 Hz, 2H), 7.71 (d, J=8.8 Hz, 2H), 7.19 (d, J=8.4 Hz, 1H), 7.13 (d,J=8.5 Hz, 1H), 6.79 (m, 1H), 5.20 (s, 2H), 4.53 (m, 1H), 4.24 (m, 2H),3.79 (m, 2H), 3.47 (m, 3H), 3.22 (m, 1H), 2.96 (m, 3H), 2.80 (m, 1H),2.63 (m, 1H), 2.50 (m, 2H), 2.37 (s, 3H), 1.17 (t, J=7.4 Hz, 3H)

Example 75:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxy-4-methylpicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

3-Hydroxy-4-methylpicolinic acid (74.9 mg, 395 μmol) was suspended inDCM (4 mL) at RT. After cooling to 0° C.,1-chloro-N,N,2-trimethylprop-1-en-1-amine (62.0 μL, 470 μmol) was addeddropwise. The RM was stirred at RT for 2 hours, then it was cooled to 0°C. again. DIPEA (164 μL, 941 μmol) was added. This solution was addeddropwise at 0° C. to a solution of2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide(Intermediate AS) (100 mg, 188 μmol) in DCM (4 mL). The RM was stirredat RT for 18 hours. The RM was diluted with DCM, washed with aq satNH₄Cl, dried over Na₂SO₄ and concentrated. The resulting solid wasfiltered off, washed with DCM and TBME and dried to give the titlecompound as a grey solid.

LC-MS: Rt=1.12 min; MS m/z [M+H]⁺ 667.2, m/z [M−H]⁻ 665.1; UPLC-MS 4

¹H NMR (400 MHz, DMSO-d₆) δ 10.88 (s, 1H), 10.37 (s, 1H), 8.00 (d, J=4.6Hz, 1H), 7.78 (d, J=8.6 Hz, 2H), 7.71 (d, J=8.8 Hz, 2H), 7.26 (d, J=4.6Hz, 1H), 6.79 (m, 1H), 5.20 (s, 2H), 4.58 (m, 1H), 4.24 (m, 2H), 4.08(m, 1H), 3.79 (m, 2H), 3.52 (m, 2H), 3.30 (m, 1H), 2.99 (m, 3H), 2.80(m, 1H), 2.67 (m, 1H), 2.51 (m, 2H), 2.24 (s, 3H), 1.18 (t, J=7.4 Hz,3H)

Example 76:2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-2-(pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To the stirred solution of2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AT) (170 mg, 314 μmol) in DMF (2 mL), the mixture of3-hydroxypicolinic acid (87.0 mg, 629 μmol), EDC.HCl (121 mg, 629 μmol),HOBt (85.0 mg, 629 μmol) and DIPEA (330 μL, 1.89 mmol) was added at RT.The RM was stirred at RT for 4 hours. The RM was concentrated underreduced pressure, water was added and the precipitate was filtered offand dried under reduced pressure. The crude product was purified byreverse phase preparative HPLC (RP-HPLC acidic 7: 30 to 40% B in 2 min,40 to 60% B in 8 min) to give the title compound.

LC-MS: Rt=0.95 min; MS m/z [M+H]⁺ 662.3, MS m/z [M−H]⁻ 660.3; UPLC-MS 1

LC-MS: Rt=4.74 min; MS m/z [M+H]⁺ 662.2, MS m/z [M−H]⁻ 660.3; UPLC-MS 2

¹H NMR (600 MHz, DMSO-d₆) δ 10.38 (s, 1H), 10.05 (s, 1H), 9.22 (m, 1H),8.67 (m, 1H), 8.39 (m, 1H), 8.03 (m, 1H), 7.68 (d, J=8.5 Hz, 1H), 7.59(m, 1H), 7.54 (m, 1H), 7.49 (d, J=8.9 Hz, 1H), 7.25 (m, 2H), 5.30 (s,2H), 4.52 (m, 1H), 3.46 (m, 2H), 3.37 (m, 1H), 3.20 (m, 1H), 2.96 (m,3H), 2.79 (m, 1H), 2.61 (m, 1H), 2.33 (s, 3H), 1.18 (t, J=7.5 Hz, 3H)

Example 77:2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(6-methylpyridin-3-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To a solution of2-(5-ethyl-2-(6-methylpyridin-3-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AU) (188 mg, 183 μmol) in DCM (1.2 mL) under argon at 0°C. was added 3-hydroxypicolinoyl chloride (Intermediate CV) (43.1 mg,274 μmol). DIPEA (159 μL, 913 μmol) was then added dropwise. Theresulting solution was stirred at 0° C. for 15 minutes, then at RT for3.5 hours. 3-Hydroxypicolinoyl chloride (Intermediate CV) (43.1 mg, 274μmol) was added to the RM at 0° C., and the RM was stirred at RT for 19hours. Then DIPEA (159 μL, 913 μmol) was added, and the RM was stirredat RT for 3.5 hours. The RM was diluted with DCM and washed with water.The aqueous layer was extracted twice with DCM. The combined organiclayers were dried through a phase separator and concentrated underreduced pressure. The crude product was purified by reverse phasepreparative HPLC (RP-HPLC acidic 1: 8 to 38% B in 20 min with a plateauat 38% for 1 min). The product containing fractions were combined andconcentrated to remove ACN and TFA. The resulting aqueous residue wasbasified with solid NaHCO₃. A precipitate was formed which was filteredoff, washed with water until neutral and dried under reduced pressure togive the title compound as a colorless solid.

LC-MS: Rt=0.99 min; MS m/z [M+H]⁺ 676.5, m/z [M−H]⁻ 674.5; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.49 (s, br, 1H), 10.08 (s, br, 1H), 9.14(d, J=2.0 Hz, 1H), 8.32 (dd, J=2.2 Hz, 8.0 Hz, 1H), 8.06 (m, 1H), 7.72(d, J=8.4 Hz, 1H), 7.63 (m, 1H), 7.53 (d, J=8.4 Hz, 1H), 7.44 (d, J=8.1Hz, 1H), 7.29 (m, 2H), 5.33 (s, 2H), 4.57 (m, 1H), 3.48 (m, 3H), 3.23(m, 1H), 3.03 (m, 3H), 2.83 (m, 1H), 2.67 (m, 1H), 2.55 (s, 3H), 2.38(s, 3H), 1.23 (t, J=7.4 Hz, 3H)

Example 78:2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To the stirred solution of 4-chloro-3-hydroxypicolinic acid (86.0 mg,493 μmol) in DMF (10 mL) at RT were added EDC.HCl (95.0 mg, 493 μmol),HOBt (66.6 mg, 493 μmol), pyridine (80.0 μL, 986 μmol) and then2-(2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AV) (180 mg, 329 μmol), and the RM was stirred at RT for14 hours. Water (20 mL) was added to the RM and it was extracted with10% MeOH in DCM (2×20 mL). The organic layers were washed with brine (20mL), dried over Na₂SO₄ and concentrated. The crude product was purifiedby column chromatography (Silica gel column: Silica 40 g, eluentDCM:MeOH 100:0 to 94:6) to give the title compound.

LC-MS: Rt=1.04 min; MS m/z [M+H]⁺ 703.6, m/z [M−H]⁻ 701.3; UPLC-MS 1

LC-MS: Rt=5.06 min; MS m/z [M+H]⁺ 703.2, m/z [M−H]⁻ 701.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.83 (s, br, 1H), 9.99 (s, 1H), 8.06 (d,J=5.0 Hz, 1H), 7.72 (d, J=8.4 Hz, 1H), 7.63 (m, 1H), 7.54 (m, 2H), 7.09(s, 1H), 5.21 (s, 2H), 4.54 (m, 1H), 4.19 (m, 4H), 3.51 (m, 3H), 3.23(m, 1H), 2.99 (m, 3H), 2.81 (m, 1H), 2.65 (m, 1H), 2.35 (s, 3H), 1.19(t, J=7.4 Hz, 3H)

Example 79:2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

4-Chloro-3-hydroxypicolinic acid (1.40 g, 8.06 mmol) was suspended inDCM (40 mL). Then 1-chloro-N,N,2-trimethylprop-1-en-1-amine (2.13 mL,16.1 mmol) was added. The RM was stirred at RT for 45 minutes.2-(2-(5,6-Dihydro-1,4-dioxin-2-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AW) (1.22 g, 1.61 mmol) was added, followed by DIPEA (5.63mL, 32.2 mmol). The RM was stirred at RT for 1 hour. Water (10 mL), aqsat NaHCO₃ (10 mL) and DCM (10 mL) were added. The aqueous layer wasextracted twice with DCM (2×10 mL). The combined organic layers weredried through a phase separator and concentrated under reduced pressure.The crude product was purified by column chromatography (RediSep Column:Silica 80 g, eluent DCM:DCM/MeOH (9/1) 100:0 to 0:100). The productcontaining fractions were combined, concentrated under vacuum and driedunder HV to give product which was not pure. This material was purifiedin 2 portions by reverse phase preparative HPLC (RP-HPLC acidic 1: 10 to100% B in 20 min and RP-HPLC acidic 1: 15 to 100% in 20 min). Theproduct containing fractions were combined, basified with aq sat NaHCO₃,extracted twice with DCM, dried through a phase separator andconcentrated under reduced pressure to give the title compound.

LC-MS: Rt=1.07 min; MS m/z [M+H]⁺ 721.3, m/z [M−H]⁻ 719.4; UPLC-MS 1

¹H NMR (600 MHz, DMSO-d₆) δ 10.83 (s, 1H), 10.19 (s, 1H), 8.06 (d, J=5.0Hz, 1H), 7.59 (m, 2H), 7.55 (d, J=5.0 Hz, 1H), 7.08 (s, 1H), 5.22 (s,2H), 4.54 (m, 1H), 4.19 (m, 4H), 3.52 (m, 3H), 3.24 (m, 1H), 2.98 (m,3H), 2.81 (m, 1H), 2.62 (m, 1H), 2.24 (s, 3H), 1.18 (t, J=7.5 Hz, 3H)

Example 80:rac-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

3-Hydroxypicolinic acid (51.3 mg, 369 μmol) was dissolved in DMF (2 mL)and then EDC.HCl (70.7 mg, 369 μmol), DIPEA (258 μL, 1.48 mmol), HOBt(49.8 mg, 369 μmol) andrac-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide.HCl(Intermediate AX) (150 mg, 246 μmol) were added at 0° C. and the RM wasstirred at RT for 16 hours. The RM was diluted with water, extractedwith EtOAc, washed with aq sat NaHCO₃ and brine and the combined organiclayers were dried over Na₂SO₄, filtered and concentrated under reducedpressure. The crude product was purified by column chromatography(Silica gel column: Silica 12 g, eluent DCM:MeOH 100:0 to 95:5) to givethe title compound as a fluffy white solid.

LC-MS: Rt=1.04 min; MS m/z [M+H]⁺ 695.1, m/z [M−H]⁻ 693.3; UPLC-MS 1

LC-MS: Rt=5.18 min; MS m/z [M+H]⁺ 695.2, m/z [M−H]⁻ 693.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.38 (s, 1H), 9.99 (s, 1H), 8.06 (m, 1H),7.72 (d, J=8.2 Hz, 1H), 7.62 (m, 1H), 7.53 (d, J=8.4 Hz, 1H), 7.28 (m,2H), 6.73 (m, 1H), 5.23 (s, 2H), 4.54 (m, 1H), 3.45 (m, 4H), 3.28 (s,3H), 3.22 (m, 1H), 2.97 (m, 3H), 2.79 (m, 1H), 2.60 (m, 3H), 2.35 (s,3H), 2.33 (m, 1H), 2.15 (m, 1H), 1.95 (m, 1H), 1.67 (m, 1H), 1.19 (t,J=7.3 Hz, 3H)

Example 80a:((R)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide)or((S)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide)and Example 80b:((R)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide)or((S)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide)

Chiral separation ofrac-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide:

Preparative chiral HPLC (instrument: Waters Prep SFC100 Mass directed;column: Chiralpak IB-N 250×30 mm 5 μm; eluent: A: 30% [MeOH+0.1% NH₃];B: 70% scCO₂; flow rate: 80 mL/min; detection: 190-400 nm; injectionvolume: 1.2 mL; Gradient: isocratic: A(30):B(70) in 17 minutes).

Example 80a: First eluting stereoisomer, white solid.

Chiral HPLC (C-HPLC 8): Rt=2.19 min, 99.5% ee

LC-MS: Rt=1.04 min; MS m/z [M+H]⁺ 695.5, m/z [M−H]⁻ 693.5; UPLC-MS 1

LC-MS: Rt=5.16 min; MS m/z [M+H]⁺ 695.3, m/z [M−H]⁻ 693.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.37 (s, br, 1H), 9.99 (s, 1H), 8.06 (m,1H), 7.72 (d, J=8.4 Hz, 1H), 7.62 (m, 1H), 7.53 (d, J=8.4 Hz, 1H), 7.29(m, 2H), 6.73 (m, 1H), 5.23 (s, 2H), 4.54 (m, 1H), 3.47 (m, 4H), 3.28(s, 3H), 3.22 (m, 1H), 2.98 (m, 3H), 2.79 (m, 1H), 2.60 (m, 3H), 2.35(s, 3H), 2.33 (m, 1H), 2.16 (m, 1H), 1.95 (m, 1H), 1.67 (m, 1H), 1.19(t, J=7.4 Hz, 3H)

Example 80b: Second eluting stereoisomer, white solid.

Chiral HPLC (C-HPLC 8): Rt=2.73 min, 98.5% ee

LC-MS: Rt=1.04 min; MS m/z [M+H]⁺ 695.5, m/z [M−H]⁻ 693.5; UPLC-MS 1

LC-MS: Rt=5.16 min; MS m/z [M+H]⁺ 695.3, m/z [M−H]⁻ 693.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.37 (s, br, 1H), 9.99 (s, 1H), 8.06 (m,1H), 7.72 (d, J=8.4 Hz, 1H), 7.62 (m, 1H), 7.52 (d, J=8.1 Hz, 1H), 7.28(m, 2H), 6.73 (m, 1H), 5.23 (s, 2H), 4.54 (m, 1H), 3.46 (m, 4H), 3.28(s, 3H), 3.22 (m, 1H), 2.97 (m, 3H), 2.79 (m, 1H), 2.61 (m, 2H), 2.41(m, 2H), 2.35 (s, 3H), 2.33 (m, 1H), 2.15 (m, 1H), 1.96 (m, 1H), 1.68(m, 1H), 1.19 (t, J=7.4 Hz, 3H)

Example 81:rac-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

3-Hydroxypicolinic acid (67.6 mg, 486 μmol) was dissolved in DCM (1 mL)and 1-chloro-N,N,2-trimethylprop-1-en-1-amine (113 μL, 851 μmol) wasadded. The RM was stirred at RT for 1 hour. Thenrac-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide(Intermediate AY) (200 mg, 243 μmol) and DIPEA (170 μL, 973 μmol) wereadded and the RM was stirred at RT for 3 hours. Water, aq sat NaHCO₃,brine and DCM were added. The aqueous layer was extracted twice withDCM. The combined organic layers were dried through a phase separatorand concentrated under reduced pressure. The crude product was purifiedby reverse phase preparative HPLC (RP-HPLC acidic 1: 10 to 100% B in 30min). The product containing fractions were combined, basified with aqsat NaHCO₃, extracted twice with DCM, dried through a phase separatorand concentrated under reduced pressure. The resulting solid waspurified by reverse phase preparative HPLC (RP-HPLC acidic 1: 25 to 75%B in 20 min with a plateau at 75% for 1 min). The product containingfractions were combined, basified with aq sat NaHCO₃, extracted twicewith DCM, dried through a phase separator and concentrated under reducedpressure to give the title compound.

LC-MS: Rt=0.97 min; MS m/z [M+H]⁺ 681.4, m/z [M−H]⁻ 679.5; UPLC-MS 4

Example 81a:((R)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamideor((S)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamideand Example 81b:((R)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamideor((S)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

Chiral separation ofrac-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide:

Preparative chiral HPLC (instrument: Waters Prep SFC100 Mass directed;column: Chiralpak IBN 250×30 mm 5 μm; eluent: A: 35% [MeOH+0.1% NH₃]; B:65% scCO₂; flow rate: 80 mL/min; detection: 190-400 nm; injectionvolume: 4 mL; gradient: isocratic: A(35):B(65) in 17 min). The peakswere dissolved in a large amount of DCM and MeOH and filtered through aPL-HCO₃ MP SPE cartridge (100 mg per 6 mL). The filtrate wasconcentrated under reduced pressure and dried under HV.

Example 81a: First eluting stereoisomer, beige solid.

Chiral HPLC (C-HPLC 8): Rt=2.51 min, 99% ee

LC-MS: Rt=1.04 min; MS m/z [M+H]⁺ 681.4, m/z [M−H]⁻ 679.4; UPLC-MS 4

LC-MS: Rt=5.23 min; MS m/z [M+H]⁺ 681.3, m/z [M−H]⁻ 679.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.85 (s, 1H), 10.38 (s, 1H), 8.06 (m, 1H),7.78 (d, J=8.4 Hz, 2H), 7.70 (d, J=8.6 Hz, 2H), 7.29 (m, 2H), 6.69 (m,1H), 5.18 (s, 2H), 4.55 (m, 1H), 3.46 (m, 4H), 3.27 (s, 3H), 3.21 (m,1H), 2.97 (m, 3H), 2.80 (m, 1H), 2.62 (m, 2H), 2.42 (m, 2H), 2.13 (m,1H), 1.93 (m, 1H), 1.66 (m, 1H), 1.17 (t, J=7.2 Hz, 3H)

Example 81b: Second eluting stereoisomer, beige solid.

Chiral HPLC (C-HPLC 8): Rt=3.23 min, 95% ee

LC-MS: Rt=1.04 min; MS m/z [M+H]⁺ 681.4, m/z [M−H]⁻ 679.4; UPLC-MS 4

Example 82:2-(4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)piperazin-1-yl)aceticacid

Step 1: ethyl2-(4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)piperazin-1-yl)acetate

Ethyl2-(4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)piperazin-1-yl)acetate(Intermediate AZ) (300 mg, 459 μmol) was dissolved in DCM (1 mL) andDIPEA (481 μL, 2.75 mmol) was added at 0° C., followed by3-hydroxypicolinoyl chloride (Intermediate CV) (217 mg, 1.38 mmol). TheRM was allowed to warm to RT and stirred at RT for 16 hours. The RM wasconcentrated under reduced pressure and washed with Et₂O to give thetitle compound.

LC-MS: Rt=1.16 min; MS m/z [M+H]⁺ 775.3; UPLC-MS 13

Step 2:2-(4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)piperazin-1-yl)aceticacid

Ethyl2-(4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)piperazin-1-yl)acetate(300 mg, 387 μmol) was dissolved in EtOH (1 mL) and water (1 mL), andLiOH.H₂O (48.7 mg, 1.16 mmol) was added and the RM was stirred at RT for2 hours. The EtOH was removed under reduced pressure and the residue wasacidified with citric acid and extracted with EtOAc. The crude productwas purified by reverse phase preparative HPLC (RP-HPLC acidic 9: 25 to35% B in 2 min, 35 to 45% B in 10 min) to give the title compound.

LC-MS: Rt=0.78 min; MS m/z [M+H]⁺ 747.2, m/z [M−H]⁻ 745.3; UPLC-MS 1

LC-MS: Rt=3.90 min; MS m/z [M+H]⁺ 747.2, m/z [M−H]⁻ 745.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.32 (s, br, 1H), 8.05 (m, 2H), 7.96 (m,1H), 7.72 (d, J=8.6 Hz, 1H), 7.28 (m, 2H), 5.20 (s, 2H), 4.53 (m, 1H),3.55-3.20 (m, 8H), 3.16 (s, 2H), 2.92 (m, 3H), 2.75 (m, 1H), 2.59 (m,5H), 1.15 (t, J=7.4 Hz, 3H)

Example 83:2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

2-(2-(3,4-Dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate BA) (190 mg, 261 μmol) was dissolved in DCM (5 mL) at 0°C. under argon. 3-Hydroxypicolinoyl chloride (Intermediate CV) (61.7 mg,392 μmol) was added, followed by slow addition of DIPEA (228 μL, 1.31mmol) over 5 minutes at 0° C. The RM was stirred at 0° C. for 20minutes, then at RT for 1.2 hours. 3-Hydroxypicolinoyl chloride(Intermediate CV) (26.0 mg, 165 μmol) and DIPEA (70.0 μL, 402 μmol) wereadded and the RM was stirred at RT for 1.5 hours. The RM was quenchedwith water (5 mL) and aq sat NaHCO₃ (5 mL), and it was extracted withDCM (4×50 mL). The combined organic layers were washed with aq satNaHCO₃ and water, dried through a phase separator and concentrated underreduced pressure. The residue was adsorbed onto Isolute and purified bycolumn chromatography (RediSep Column: Silica 24 g, eluent DCM:MeOH100:0 to 90:10). The product containing fractions were combined andconcentrated under reduced pressure. This material was adsorbed ontoIsolute and purified by column chromatography (RediSep Column: Silica 24g, eluent DCM:MeOH 100:0 to 95:5). The resulting solid was furtherpurified by SFC (SFC 2). The product containing fractions were combined,concentrated under reduced pressure and dried under HV to give the titlecompound as a slightly yellow solid.

LC-MS: Rt=1.02 min; MS m/z [M+H]⁺ 667.4, m/z [M−H]⁻ 665.4; UPLC-MS 3

¹H NMR (600 MHz, DMSO-d₆) δ 10.00 (s, br, 2H), 8.07 (m, 1H), 7.74 (d,J=8.4 Hz, 1H), 7.63 (m, 1H), 7.54 (d, J=8.3 Hz, 1H), 7.29 (m, 2H), 5.87(m, 1H), 5.24 (s, 2H), 4.55 (m, 1H), 4.11 (m, 2H), 3.46 (m, 3H), 3.22(m, 1H), 2.98 (m, 3H), 2.80 (m, 1H), 2.62 (m, 1H), 2.36 (s, 3H), 2.19(m, 2H), 1.85 (m, 2H), 1.19 (t, J=7.4 Hz, 3H)

Example 84:2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

3-Hydroxypicolinic acid (83.0 mg, 600 μmol) was dissolved in DCM (2 mL)and 1-chloro-N,N,2-trimethylprop-1-en-1-amine (139 μL, 1.05 mmol) wasadded. The RM was stirred at RT for 2 hours. Then2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide(Intermediate BB) (159 mg, 300 μmol) was added, followed by DIPEA (262μL, 1.50 mmol). The RM was stirred at RT for 1 day. Water, aq sat NaHCO₃and DCM were added. The aqueous layer was extracted twice with DCM. Thecombined organic layers were dried through a phase separator andconcentrated under reduced pressure. The crude product was purified byreverse phase preparative HPLC (RP-HPLC acidic 1: 25 to 75% B in 30min). The product containing fractions were combined, basified with aqsat NaHCO₃, extracted twice with DCM, dried through a phase separatorand concentrated under reduced pressure. The resulting solid waspurified by SFC (SFC 4) to give the title compound.

LC-MS: Rt=1.02 min; MS m/z [M+H]⁺ 653.6, m/z [M−H]⁻ 651.4; UPLC-MS 1

LC-MS: Rt=5.04 min; MS m/z [M+H]⁺ 653.3, m/z [M−H]⁻ 651.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.85 (s, 1H), 10.38 (s, br, 1H), 8.06 (m,1H), 7.78 (d, J=8.6 Hz, 2H), 7.70 (d, J=8.7 Hz, 2H), 7.29 (m, 2H), 5.83(m, 1H), 5.18 (m, 2H), 4.55 (m, 1H), 4.08 (m, 2H), 3.44 (m, 3H), 3.24(m, 1H), 2.98 (m, 3H), 2.80 (m, 1H), 2.62 (m, 1H), 2.16 (m, 2H), 1.83(m, 2H), 1.16 (t, J=7.4 Hz, 3H)

Example 85:(S)-2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To the stirred solution of(S)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide.HCl(Intermediate BC) (550 mg, 915 μmol) in DMF (6 mL) were added4-chloro-3-hydroxypicolinic acid (318 mg, 1.83 mmol), EDC.HCl (351 mg,1.83 mmol), HOBt (247 mg, 1.83 mmol) and DIPEA (959 μL, 5.49 mmol) atRT. The RM was stirred at RT for 16 hours. The RM was concentrated underreduced pressure, water was added and the precipitate was filtered offand dried under reduced pressure. The crude product was purified byreverse phase preparative HPLC (RP-HPLC acidic 4: 20 to 30% B in 2 min,30 to 65% B in 8 min) to give the title compound.

LC-MS: Rt=1.11 min; MS m/z [M+H]⁺ 720.3, m/z [M−H]⁻ 718.3; UPLC-MS 1

LC-MS: Rt=5.52 min; MS m/z [M+H]⁺ 720.2, m/z [M−H]⁻ 718.2; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.87 (s, br, 1H), 10.00 (s, 1H), 8.02 (m,1H), 7.69 (d, J=8.4 Hz, 1H), 7.63 (m, 1H), 7.53 (m, 2H), 5.13 (s, 2H),4.75 (m, 1H), 4.52 (m, 1H), 3.74 (m, 1H), 3.55 (m, 5H), 3.22 (m, 2H),2.95 (m, 3H), 2.77 (m, 1H), 2.59 (m, 1H), 2.34 (s, 3H), 1.83 (m, 3H),1.53 (m, 1H), 1.17 (d, J=7.1 Hz, 3H)

Example 86:(R)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:(R)-2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-3-methylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

(R)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate BE) (350 mg, 603 μmol),5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (Intermediate CY)(155 mg, 634 μmol) and HATU (241 mg, 634 μmol) were suspended in DCM (10mL) and DMF (200 μL) and the mixture was cooled to 0° C. DIPEA (263 μL,1.51 mmol) was added, and the suspension was stirred at 0° C. for 10minutes and at RT for 30 minutes. Water (10 mL), aq sat NaHCO₃ (10 mL)and DCM (10 mL) were added. The aqueous layer was extracted with DCM(2×10 mL), and the organic layers were combined, dried through a phaseseparator and concentrated under reduced pressure. The crude product waspurified by column chromatography (RediSep Column: Silica 24 g, eluentDCM:MeOH 100:0 to 75:25). The product containing fractions werecombined, concentrated under vacuum and dried under HV to afford thetitle compound as a brown oil. The product was heated in water:MeOH(1:3) until fully dissolved and left to cool to RT. The suspension wasfiltered off and washed with MeOH, then dried to give the title compoundas beige solid.

LC-MS: Rt=1.23 min; MS m/z [M+H]⁺ 806.5, m/z [M−H]⁻ 804.5; UPLC-MS 1

Step 2:(R)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

(R)-2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-3-methylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(598 mg, 600 μmol) was dissolved in DCM (1 mL), and TFA (1.00 mL, 13.0mmol) was added. The RM was stirred at 45° C. for 1 day. The RM wasconcentrated under reduced pressure, and water (10 mL), aq sat NaHCO₃(10 mL) and DCM (10 mL) were added. The aqueous layer was extracted withDCM (2×10 mL). The combined organic layers were dried through a phaseseparator and concentrated under reduced pressure. The crude product waspurified by column chromatography (RediSep Column: Silica 40 g, eluentDCM:MeOH 100:0 to 95:05). The product containing fractions werecombined, concentrated under vacuum and dried under HV. This materialwas further purified in 2 portions by reverse phase preparative HPLC(RP-HPLC acidic 1: 20 to 80% B in 20 min with a plateau at 80% for 1 minand RP-HPLC acidic 1: 25 to 75% B in 20 min with a plateau at 75% for 1min). All product containing fractions were combined, basified with aqsat NaHCO₃, extracted twice with DCM, dried through a phase separatorand concentrated under reduced pressure. The concentrated fractions weresuspended in MeOH, sonicated and filtered. The cake was dried under HV.This material was heated in water and MeOH (1:3) until fully dissolvedand left to cool. The suspension was filtered off and washed with MeOH,then dried to give the title compound as beige solid characterized bythe XRPD diffractogram in FIG. 2 The table below shows the mostprominent peaks (deg 2theta) of the XRPD diffractogram of FIG. 2 .

The sodium salt was prepared analogous to the general procedure.

LC-MS: Rt=1.06 min; MS m/z [M+H]⁺ 716.1, m/z [M−H]⁻ 714.4; UPLC-MS 1

LC-MS: Rt=5.26 min; MS m/z [M+H]⁺ 716.1, m/z [M−H]⁻ 714.5; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.35 (s, 1H), 10.15 (d, J=19.4 Hz, 1H),8.57 (d, J=3.7 Hz, 1H), 8.06 (d, J=8.6 Hz, 1H), 7.96 (m, 1H), 7.71 (d,J=8.6 Hz, 1H), 6.83 (m, 1H), 5.32 (s, 2H), 4.82-4.36 (m, 1H), 4.25 (m,2H), 3.80 (m, 2H), 3.71 (m, 2H), 3.50 (m, 1H), 3.16 (m, 2H), 2.92 (m,1H), 2.81 (m, 1H), 2.66 (m, 1H), 2.50 (m, 2H), 2.44 (s, 3H), 1.40 (m,3H), 1.20 (t, J=7.4 Hz, 3H)

Angle d Value 2-Theta° Angstrom Intensity 10.29 8.59 high 13.31 6.65medium 14.01 6.32 low 15.26 5.80 medium 17.34 5.11 medium

Example 87:(S)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:(S)-2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2-methylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

(S)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(2-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate BH) (58.0 mg, 80.0 μmol) and5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (Intermediate CY)(19.5 mg, 80.0 μmol) were mixed in DMF (1 mL) at RT under argon. HATU(36.5 mg, 96.0 μmol) was added, followed by DIPEA (69.9 μL, 400 μmol).The RM was stirred at RT for 1.75 hours. Water (10 mL) was added and theRM was extracted with EtOAc (4×50 mL). The organic layers were combined,washed with water (2×10 mL) and brine (2×10 mL), dried trough a phaseseparator and concentrated under reduced pressure. The crude product waspurified by column chromatography (RediSep Column: Silica 24 g, eluentDCM:DCM/MeOH (8/2) 100:0 to 60:40). The product containing fractionswere combined and concentrated to give the title compound as a beigesolid.

LC-MS: Rt=1.21 min; MS m/z [M+H]⁺ 806.3, m/z [M−H]⁻ 804.3; UPLC-MS 1

Step 2:(S)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

(S)-2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2-methylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(33.0 mg, 41.0 μmol) in TFA (1.00 mL, 13.0 mmol) was stirred at 50° C.for 3 hours and then at RT overnight. The RM was concentrated and driedunder HV, water (10 mL) was added, and the residue was extracted withDCM (4×15 mL). The organic layers were washed with aq sat NaHCO₃ (5 mL)and brine (5 mL), dried through a phase separator and concentrated underreduced pressure. The crude product was adsorbed onto Isolute andpurified by column chromatography (RediSep Column: Silica 12 g, eluentDCM:DCM/MeOH (8/2) 100:0 to 70:30). The product containing fractionswere combined and concentrated under reduced pressure to give the titlecompound as a beige solid.

LC-MS: Rt=1.07 min; MS m/z [M+H]⁺ 716.3, m/z [M−H]⁻ 714.3; UPLC-MS 1

LC-MS: Rt=5.35 min; MS m/z [M+H]⁺ 716.2, m/z [M−H]⁻ 714.3; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.35 (s, br, 2H), 8.55 (d, J=5.3 Hz, 1H),8.05 (d, J=8.5 Hz, 1H), 7.96 (m, 1H), 7.71 (d, J=8.9 Hz, 1H), 6.83 (m,1H), 5.30 (m, 2H), 4.50 (m, 1H), 4.25 (m, 2H), 3.80 (m, 2H), 3.62 (m,1H), 3.45 (m, 2H), 2.98 (m, 1H), 2.81 (m, 3H), 2.61 (m, 1H), 2.50 (m,2H), 2.43 (d, J=4.4 Hz, 3H), 1.24 (s, 3H), 1.17 (m, 3H)

Example 88:(R)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:(R)-2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2-methylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

(R)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(2-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate BJ) (93.0 mg, 160 μmol) and5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (Intermediate CY)were mixed in DMF (2 mL) at RT under argon. HATU (75.0 mg, 192 μmol) andDIPEA (140 μL, 802 μmol) were added. The RM was stirred at RT for 40minutes, then it was quenched with water (5 mL) and extracted with EtOAc(3×50 mL). The organic layers were washed with water (2×10 mL) and brine(2×10 mL), dried through a phase separator and concentrated underreduced pressure. The crude product was purified by reverse phasepreparative HPLC (RP-HPLC acidic 1: 15 to 85% B in 20 min with a plateauat 85% for 1 min). The product containing fractions were combined andlyophilized to give the title compound as a white solid.

LC-MS: Rt=1.21 min; MS m/z [M+H]⁺ 806.3, m/z [M−H]⁻ 804.4; UPLC-MS 1

Step 2:(R)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

(R)-2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2-methylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(70.7 mg, 88.0 μmol) was mixed with TFA (2.00 mL, 26.0 mmol) and stirredat 50° C. for 2.75 hours. The RM was concentrated under reducedpressure, dried under HV and extracted with DCM (4×30 mL). The organiclayers were washed with aq sat NaHCO₃ (15 mL) and brine (20 mL), driedthrough a phase separator and concentrated under reduced pressure. Thecrude product was adsorbed onto Isolute and purified by columnchromatography (RediSep Column: Silica 12 g, eluent DCM:MeOH 100:0 to80:20). The product containing fractions were combined and concentratedto give the title compound as a colorless solid.

LC-MS: Rt=1.07 min; MS m/z [M+H]⁺ 716.3, m/z [M−H]⁻ 714.3; UPLC-MS 1

LC-MS: Rt=5.33 min; MS m/z [M+H]⁺ 716.2, m/z [M−H]⁻ 714.2; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.36 (s, 1H), 10.22 (d, J=12.6 Hz, 1H),8.57 (d, J=4.5 Hz, 1H), 8.05 (d, J=8.6 Hz, 1H), 7.96 (m, 1H), 7.71 (d,J=8.1 Hz, 1H), 6.83 (m, 1H), 5.43-5.20 (m, 2H), 4.51 (m, 1H), 4.25 (m,2H), 3.80 (m, 2H), 3.63 (m, 1H), 3.46 (m, 2H), 3.26 (m, 1H), 2.99 (m,1H), 2.81 (m, 2H), 2.60 (m, 1H), 2.50 (m, 2H), 2.44 (d, J=4.1 Hz, 3H),1.18 (m, 3H), 0.92-0.67 (m, 3H)

Example 89:(R)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-(4-(3-hydroxypicolinoyl)-3-methylpiperazin-1-yl)-5-methyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

To a suspension of 3-hydroxypicolinic acid (47.1 mg, 339 μmol) in DCM (3mL) was added 1-chloro-N,N,2-trimethylprop-1-en-1-amine (49.0 μL, 373μmol) and the RM was stirred at RT for 40 minutes. DIPEA (148 μL, 847μmol) was added slowly, followed by(R)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide(Intermediate BL) (100 mg, 169 μmol). The RM was stirred at RT for 16.5hours. To a suspension of 3-hydroxypicolinic acid (23.6 mg, 169 μmol) inDCM (2 mL) was added 1-chloro-N,N,2-trimethylprop-1-en-1-amine (25.0 μL,187 μmol) and the RM was stirred at RT for 2 hours. The first RM wasadded dropwise and the resulting mixture was stirred at RT for 2.5hours. DIPEA (74.0 μL, 424 μmol) was added slowly, and the RM wasstirred at RT for 17 hours. The RM was concentrated to dryness, and theresidue was purified by column chromatography (RediSep Column: Silica 4g, eluent DCM:DCM/MeOH (9/1) 100:0 to 60:40). The product containingfractions were combined and concentrated under reduced pressure. Thismaterial was purified in 2 portions by reverse phase preparative HPLC(2×RP-HPLC acidic 1: 18 to 48% B in 20 min). The product containingfractions were combined, basified with solid NaHCO₃, sonicated andconcentrated to remove the ACN/TFA. The residue was extracted twice withDCM and a small amount of MeOH. The combined organic layers were driedthrough a phase separator and concentrated under reduced pressure togive the title compound.

LC-MS: Rt=1.01 min; MS m/z [M+H]⁺ 653.2, m/z [M−H]⁻ 651.0; UPLC-MS 4

¹H NMR (600 MHz, DMSO-d₆) δ 10.93 (s, 1H), 10.33 (s, br, 1H), 8.05 (m,1H), 7.79 (d, J=8.0 Hz, 2H), 7.71 (d, J=7.9 Hz, 2H), 7.28 (m, 2H), 6.81(m, 1H), 5.23 (s, 2H), 4.83-4.36 (m, 1H), 4.24 (m, 2H), 3.79 (m, 2H),3.64 (m, 1H), 3.45 (m, 1H), 3.32 (m, 1H), 3.19 (m, 1H), 2.80 (m, 1H),2.64 (m, 1H), 2.60 (s, 3H), 2.51 (m, 2H), 1.38 (m, 3H)

Example 90:2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-(4-(3-hydroxy-4-methylpicolinoyl)piperazin-1-yl)-5-methyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

3-Hydroxy-4-methylpicolinic acid (73.3 mg, 386 μmol) was suspended inDCM (4 mL) at RT. The mixture was cooled to 0° C. and1-chloro-N,N,2-trimethylprop-1-en-1-amine (64.0 μL, 483 μmol) was addeddropwise. The RM was stirred at RT for 2 hours, then it was cooled to 0°C. again. DIPEA (169 μL, 966 μmol) was added. This solution was addeddropwise at 0° C. to a solution of2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide(Intermediate BN) (100 mg, 193 μmol) in DCM (4 mL). The RM was stirredat RT for 18 hours. The RM was diluted with DCM, washed with aq satNH₄Cl, dried over Na₂SO₄, concentrated and dried. The crude product waspurified by reverse phase preparative HPLC (RP-HPLC acidic 1: 5 to 100%B in 20 min). The product containing fractions were combined, the ACNwas removed and the residue was lyophilized to give the title compoundas a foam.

LC-MS: Rt=1.07 min; MS m/z [M+H]⁺ 653.2, m/z [M−H]⁻ 651.1; UPLC-MS 4

¹H NMR (400 MHz, DMSO-d₆) δ 10.89 (s, 1H), 10.47 (s, br, 1H), 8.03 (d,J=4.7 Hz, 1H), 7.79 (d, J=8.6 Hz, 2H), 7.71 (d, J=8.8 Hz, 2H), 7.30 (d,J=4.8 Hz, 1H), 6.81 (m, 1H), 5.22 (s, 2H), 4.55 (m, 1H), 4.24 (m, 2H),4.04 (m, 1H), 3.79 (m, 2H), 3.50 (m, 2H), 3.31 (m, 1H), 3.03 (m, 1H),2.81 (m, 1H), 2.63 (m, 1H), 2.58 (s, 3H), 2.51 (m, 2H), 2.26 (s, 3H)

Example 91:2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-(4-(3-hydroxy-6-methylpicolinoyl)piperazin-1-yl)-5-methyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

3-Hydroxy-6-methylpicolinic acid (90.0 mg, 386 μmol) was suspended inDCM (4 mL) at RT. The mixture was cooled to 0° C. and1-chloro-N,N,2-trimethylprop-1-en-1-amine (56.0 μL, 425 μmol) was addeddropwise. The RM was stirred at RT for 2 hours, then it was cooled to 0°C. again. DIPEA (169 μL, 966 μmol) was added. This solution was addeddropwise at 0° C. to a solution of2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide(Intermediate BN) (100 mg, 193 μmol) in DCM (4 mL). The RM was stirredat RT for 18 hours. The RM was diluted with DCM, and the organic phasewas washed with aq sat NH₄Cl, dried over Na₂SO₄, concentrated and dried.The crude product was purified by reverse phase preparative HPLC(RP-HPLC acidic 1: 5 to 100% B in 20 min). The product containingfractions were combined, the ACN was removed and the residue waslyophilized to give the title compound.

LC-MS: Rt=1.02 min; MS m/z [M+H]⁺ 653.2, m/z [M−H]⁻ 651.1; UPLC-MS 4

¹H NMR (400 MHz, DMSO-d₆) δ 10.87 (s, 1H), 10.47 (s, br, 1H), 7.79 (d,J=8.6 Hz, 2H), 7.71 (d, J=8.8 Hz, 2H), 7.35 (d, J=8.5 Hz, 1H), 7.26 (d,J=8.5 Hz, 1H), 6.81 (m, 1H), 5.22 (s, 2H), 4.52 (m, 1H), 4.24 (m, 2H),3.80 (m, 2H), 3.46 (m, 3H), 3.26 (m, 1H), 3.00 (m, 1H), 2.80 (m, 1H),2.63 (m, 1H), 2.58 (s, 3H), 2.51 (m, 2H), 2.43 (s, 3H)

Example 92:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide

3-Hydroxypicolinic acid (114 mg, 801 μmol) was dissolved in DCM (5 mL)at RT under argon. 1-Chloro-N,N,2-trimethylprop-1-en-1-amine (118 mg,881 μmol) was added and the RM was stirred at RT for 1.75 hours. Asolution of2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide(Intermediate BP) (310 mg, 401 μmol) in DCM (2.2 mL) and DIPEA (350 μL,2.00 mmol) was added. The brown solution was stirred at RT for 2.5hours. The RM was quenched with water (5 mL) and aq sat NaHCO₃ (5 mL).Then it was extracted 4 times with DCM (4×20 mL). The organic layer waswashed with water and aq sat NaHCO₃, dried through a phase separator andconcentrated under reduced pressure. The residue was adsorbed ontoIsolute and purified by column chromatography (RediSep Column: Silica 40g, eluent DCM:MeOH 100:0 to 60:40). The product containing fractionswere combined and concentrated under reduced pressure. The resultingsolid was adsorbed onto Isolute and purified by column chromatography(RediSep Column: Silica 24 g, eluent DCM:MeOH 100:0 to 95:05). The crudeproduct was further purified by SFC (SFC 3) gave the title compound as awhite solid.

LC-MS: Rt=0.99 min; MS m/z [M+H]⁺ 671.4, m/z [M−H]⁻ 669.4; UPLC-MS 4

¹H NMR (600 MHz, DMSO-d₆) δ 10.70 (s, br, 1H), 10.40 (s, br, 1H), 8.23(t, J=8.1 Hz, 1H), 8.07 (m, 1H), 7.81 (dd, J=2.1 Hz, 11.1 Hz, 1H), 7.57(dd, J=2.0 Hz, 8.6 Hz, 1H), 7.29 (m, 2H), 6.81 (m, 1H), 5.30 (s, 2H),4.55 (m, 1H), 4.25 (m, 2H), 3.80 (m, 2H), 3.48 (m, 2H), 3.40 (m, 1H),3.22 (m, 1H), 2.97 (m, 3H), 2.80 (m, 1H), 2.63 (m, 1H), 2.51 (m, 2H),1.17 (t, J=7.7 Hz, 3H)

Example 93:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

Step 1:2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

2-(2-(3,6-Dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AJ) (97.0 mg, 139 μmol) and5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (Intermediate CY)(59.0 mg, 242 μmol) were dissolved in DCM (10 mL) and HATU (82.6 mg, 218μmol) was added, followed by DIPEA (111 μL, 695 μmol). The RM wasstirred at RT for 45 minutes. Water (10 mL), aq sat NaHCO₃ (10 mL) andDCM (10 mL) were added. The aqueous layer was washed twice with DCM(2×10 mL). The combined organic layers were dried through a phaseseparator and concentrated under reduced pressure. The crude product waspurified by column chromatography (RediSep Column: Silica 12 g, eluentDCM:DCM/MeOH (1/1) 100:0 to 88:12) to afford the title compound.

LC-MS: Rt=1.18 min; MS m/z [M+H]⁺ 790.4, m/z [M−H]⁻ 788.4; UPLC-MS 1

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

2-(6-(4-(5-(Benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide(131 mg, 124 μmol) was dissolved in DCM (2 mL) and BCl₃ 2M in DCM (124μL, 249 μmol) was added. The solution stayed a solution and was stirredat RT for 1 hour. The solution had turned into a suspension. Thesuspension was stirred at RT for 30 minutes. The RM was purged withargon and BCl₃ 2M in DCM (124 μL, 249 μmol) was added. The suspensionwas stirred at RT overnight. Water (10 mL), aq sat NaHCO₃ (10 mL) andDCM (10 mL) were added. The aqueous layer was washed twice with DCM(2×10 mL). The combined organic layers were dried through a phaseseparator and concentrated under reduced pressure. The crude product waspurified in 2 portions by column chromatography (RediSep Column: Silica4 g, eluent DCM:DCM/MeOH (1/1) 100:0 to 90:10) and (RediSep Column:Silica 12 g, eluent DCM:DCM/MeOH (1/1) 100:0 to 90:10). The productcontaining fractions were combined, concentrated under vacuum and driedunder HV to afford a yellow solid. The yellow solid was purified byreverse phase preparative HPLC (RP-HPLC basic 1: 5 to 95% B in 20 minwith a plateau at 95% for 1 min). The product containing fractions werecombined, basified with aq sat NaHCO₃, extracted twice with DCM, driedthrough a phase separator and concentrated under reduced pressure togive the title compound. The product was dissolved in DCM (5 mL) andEtOH (1 mL). The mixture was standing around for crystallization. Thecrystals were filtered and washed with a small amount of Et₂O. Then theywere dried under HV to give the title compound as a colourless solid.

LC-MS: Rt=1.03 min; MS m/z [M+H]⁺ 700.4, m/z [M−H]⁻ 698.3; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.21 (m, 2H), 8.58 (s, 1H), 7.58 (m, 2H),6.82 (m, 1H), 5.26 (s, 2H), 4.52 (m, 1H), 4.26 (m, 2H), 3.81 (m, 2H),3.47 (m, 3H), 3.25 (m, 1H), 2.99 (m, 3H), 2.81 (m, 1H), 2.63 (m, 1H),2.50 (m, 2H), 2.44 (s, 3H), 2.24 (s, 3H), 1.19 (t, J=7.5 Hz, 3H)

Example 94:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

2-(2-(3,6-Dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AM) (657 mg, 1.11 mmol) and 3-hydroxypicolinoyl chloride(Intermediate CV) (227 mg, 1.44 mmol) were dissolved in DCM (5 mL) andDIPEA (967 μL, 5.54 mmol) was added. The RM was stirred at RT for 30minutes. 3-Hydroxypicolinoyl chloride (Intermediate CV) (227 mg, 1.44mmol) and DIPEA (967 μL, 5.54 mmol) were added again and the RM wascontinued stirring at RT for 3.5 hours. Water (10 mL), aq sat NaHCO₃ (10mL) and DCM (10 mL) were added. The aqueous layer was washed twice withDCM (2×10 mL). The combined organic layers were dried through a phaseseparator and concentrated under reduced pressure. The crude product wassuspended in DCM and ACN and sonicated for 5 minutes. Then it wasfiltered. The cake was adsorbed onto Isolute and purified in 2 portionsby column chromatography (RediSep Column: Silica 24 g, eluentDCM:DCM/MeOH (9/1) 100:0 to 0:100) and (RediSep Column: Silica 24 g,eluent DCM:DCM/MeOH (9/1) 100:0 to 60:40) to afford the title compoundas a white solid.

LC-MS: Rt=1.03 min; MS m/z [M+H]⁺ 685.3, m/z [M−H]⁻ 683.2; UPLC-MS 3

¹H NMR (400 MHz, DMSO-d₆) δ 10.47 (s, br, 1H), 10.07 (s, 1H), 8.08 (m,1H), 7.79 (d, J=12.9 Hz, 1H), 7.67 (d, J=8.1 Hz, 1H), 7.32 (m, 2H), 6.82(m, 1H), 5.29 (s, 2H), 4.55 (m, 1H), 4.26 (m, 2H), 3.81 (m, 2H), 3.46(m, 3H), 3.23 (m, 1H), 2.97 (m, 3H), 2.81 (m, 1H), 2.63 (m, 1H), 2.53(m, 2H), 2.35 (s, 3H), 1.19 (t, J=7.3 Hz, 3H)

Example 95:2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

4-Chloro-3-hydroxypicolinic acid (148 mg, 852 μmol) was dissolved in DCM(8 mL) at RT under argon. 1-Chloro-N,N,2-trimethylprop-1-en-1-amine (127mg, 954 μmol) was added and the RM was stirred at RT for 2 hours.2-(2-(3,6-Dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AM) (240 mg, 341 μmol) and DIPEA (238 μL, 1.36 mmol) wereadded. The RM was stirred at RT for 1.5 hours. The RM was quenched withwater (10 mL) and aq sat NaHCO₃ (10 mL). Then it was extracted 4 timeswith DCM (4×50 mL). The organic layer was washed with water (10 mL),dried through a phase separator and concentrated under reduced pressure.The residue was adsorbed onto Isolute and purified by columnchromatography (RediSep Column: Silica 24 g, eluent DCM:MeOH 100:0 to75:25). The product containing fractions were combined and concentratedunder reduced pressure. The residue was purified by columnchromatography (RediSep Column: Silica 24 g, eluent DCM:MeOH 100:0 to95:5). The product containing fractions were combined and concentratedunder reduced pressure. The residue was purified by columnchromatography (RediSep Column: Silica 24 g, eluent DCM:MeOH 100:0 to95:5). The solid was purified by reverse phase preparative HPLC (RP-HPLCacidic 1: 30 to 70% B in 20 min with a plateau at 70% for 1 min). Theproduct containing fractions were combined, basified with aq sat NaHCO₃and the ACN was removed under reduced pressure. The aqueous layer wasextracted 4 times with DCM (4×50 mL), dried through a phase separatorand concentrated under reduced pressure to give the title compound as abeige solid.

LC-MS: Rt=1.12 min; MS m/z [M+H]⁺ 719.3/721.3, m/z [M−H]⁻ 717.2/719.2;UPLC-MS 1

¹H NMR (600 MHz, DMSO-d₆) δ 10.83 (s, 1H), 10.06 (s, 1H), 8.07 (d, J=5.1Hz, 1H), 7.78 (d, J=13.1 Hz, 1H), 7.67 (d, J=8.2 Hz, 1H), 7.56 (d, J=5.4Hz, 1H), 6.82 (m, 1H), 5.29 (s, 2H), 4.54 (m, 1H), 4.25 (m, 2H), 3.80(m, 2H), 3.59 (m, 1H), 3.50 (m, 2H), 3.25 (m, 1H), 2.98 (m, 3H), 2.82(m, 1H), 2.64 (m, 1H), 2.52 (m, 2H), 2.35 (s, 3H), 1.19 (d, J=7.2 Hz,3H)

Example 96:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AK) (450 mg, 604 μmol) was dissolved in DCM (6 mL) at 0°C. under argon. 3-Hydroxypicolinoyl chloride (Intermediate CV) (119 mg,755 μmol) was added, followed by slow addition of DIPEA (106 μL, 607μmol). The RM was stirred at RT for 1.75 hours. 3-Hydroxypicolinoylchloride (Intermediate CV) (125 mg, 793 μmol) and DIPEA (320 μL, 1.83mmol) were added again and the RM was stirred at RT for 2.5 hours.3-Hydroxypicolinoyl chloride (Intermediate CV) (150 mg, 952 μmol) andDIPEA (110 μL, 630 μmol) were added again. The RM was stirred at RT for1 hour. 3-Hydroxypicolinoyl chloride (Intermediate CV) (110 mg, 698μmol) was added again and the RM was stirred at RT for 2.25 hours. Thenit was stored in the freezer overnight. The RM was quenched with water(10 mL) and aq sat NaHCO₃ (10 mL). Then it was extracted 4 times withDCM (4×40 mL). The organic layer was washed with water (10 mL), aq satNaHCO₃ (10 mL) and water (15 mL), dried through a phase separator andconcentrated under reduced pressure. The residue was adsorbed ontoIsolute and purified by column chromatography (RediSep Column: Silica 24g, eluent DCM:MeOH 100:0 to 75:25). The product containing fractionswere combined and concentrated under reduced pressure. The resultingsolid was purified in 2 equal portions by reverse phase preparative HPLC(2×RP-HPLC acidic 1: 10 to 60% B in 20 min with a plateau at 60% for 1min). The product containing fractions were combined, basified with aqsat NaHCO₃ and the ACN was removed under reduced pressure. The aqueouslayer was extracted 4 times with DCM (4×50 mL). The aqueous layer wasextracted three times with EtOAc (3×60 mL). The combined organic layerswere dried through a phase separator and concentrated under reducedpressure to give the title compound as a beige solid characterized bythe XRPD diffractogram in FIG. 5 . The table below shows the mostprominent peaks (deg 2theta) of the XRPD diffractogram of FIG. 5 .

The sodium salt was prepared analogous to the general procedure.

LC-MS: Rt=1.03 min; MS m/z [M+H]⁺ 687.3/689.3, m/z [M−H]⁻ 685.2/687.1;UPLC-MS 1

¹H NMR (600 MHz, DMSO-d₆) δ 10.38 (s, 1H), 10.36 (s, 1H), 8.06 (m, 2H),7.96 (m, 1H), 7.71 (dd, J=1.9 Hz, 8.4 Hz, 1H), 7.29 (m, 2H), 6.83 (m,1H), 5.31 (s, 2H), 4.55 (m, 1H), 4.25 (m, 2H), 3.80 (m, 2H), 3.46 (m,3H), 3.22 (m, 1H), 2.97 (m, 3H), 2.80 (m, 1H), 2.62 (m, 1H), 2.52 (m,2H), 1.18 (t, J=7.5 Hz, 3H)

Angle d Value 2-Theta° Angstrom Intensity 13.20 6.70 medium 14.78 5.99medium 15.97 5.55 medium 16.91 5.24 low 19.95 4.45 medium 20.85 4.26medium 24.43 3.64 high 25.47 3.49 low 31.06 2.88 low

Example 97:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-fluoro-3-hydroxy-5-methylpicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-(4-(3-(benzyloxy)-4-fluoro-5-methylpicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

To a beige suspension ofN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AK) (65.0 mg, 109 μmol) and3-(benzyloxy)-4-fluoro-5-methylpicolinic acid (Intermediate DA) (30.0mg, 109 μmol) in DMF (1 mL) were added DIPEA (95.0 μL, 546 μmol) andHATU (49.8 mg, 131 μmol). The yellow RM was stirred at RT for 20minutes. The RM was diluted with water and extracted twice with DCM. Thecombined organic layers were dried through a phase separator andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (RediSep Column: Silica 4 g, eluent DCM:DCM/MeOH(8/2) 100:0 to 50:50). The product containing fractions were combined,concentrated under reduced pressure and dried under HV. The residue waspurified by column chromatography (RediSep Column: Silica 4 g, eluentDCM:DCM/MeOH (8/2) 100:0 to 65:35) to give the title compound as acolourless solid.

LC-MS: Rt=1.28 min; MS m/z [M+H]⁺ 809.4/811.4, m/z [M−H]⁻ 807.1/809.1;UPLC-MS 1

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-fluoro-3-hydroxy-5-methylpicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

A light yellow solution of2-(6-(4-(3-(benzyloxy)-4-fluoro-5-methylpicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(59.0 mg, 69.0 μmol) in TFA (513 μL) was stirred at 50° C. for 3 hours,then at RT overnight. The RM was diluted with DCM and washed with aq satNaHCO₃. The aqueous layer was extracted twice with DCM. The combinedorganic layers were dried through a phase separator and concentratedunder reduced pressure. The residue was adsorbed onto Isolute andpurified by column chromatography (RediSep Column: Silica 4 g, eluentDCM:DCM/MeOH (8/2) 100:0 to 65:35) to give the title compound as a beigesolid.

LC-MS: Rt=1.09 min; MS m/z [M+H]⁺ 719.2/721.2, m/z [M−H]⁻ 717.3/719.3;UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.65 (s, 1H), 10.35 (s, 1H), 8.05 (d, J=8.4Hz, 1H), 7.99 (d, J=8.4 Hz, 1H), 7.96 (m, 1H), 7.71 (dd, J=2.2 Hz, 8.8Hz, 1H), 6.83 (m, 1H), 5.31 (s, 2H), 4.53 (m, 1H), 4.25 (m, 2H), 3.80(m, 2H), 3.49 (m, 3H), 3.24 (m, 1H), 2.99 (m, 3H), 2.80 (m, 1H), 2.62(m, 1H), 2.50 (m, 2H), 2.24 (s, 3H), 1.19 (t, J=7.2 Hz, 3H)

Example 98:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxy-4-(trifluoromethoxy)picolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-methoxy-4-(trifluoromethoxy)picolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To a white suspension of 3-methoxy-4-(trifluoromethoxy)picolinic acid(Intermediate CZ) (43.6 mg, 180 μmol) in DCM (2.5 mL) was added1-chloro-N,N,2-trimethylprop-1-en-1-amine (51.7 μL, 375 μmol). The whitesuspension was stirred for at RT for 1 hour. DIPEA (79.0 μL, 451 μmol)was added slowly, followed byN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AK) (85.0 mg, 150 μmol). The light brown turbid RM wasstirred at RT for 3 hours. The RM was diluted with DCM and washed withwater. The aqueous layer was extracted with DCM. The combined organiclayers were dried through a phase separator and concentrated underreduced pressure. The crude product was purified by columnchromatography (RediSep Column: Silica 4 g, eluent DCM:DCM/MeOH (8/2)100:0 to 70:30) to give the title compound as a yellow solid.

LC-MS: Rt=1.22 min; MS m/z [M+H]⁺ 785.5/787.5, m/z [M−H]⁻ 783.4/785.4;UPLC-MS 1

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxy-4-(trifluoromethoxy)picolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

A light yellow RM ofN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-methoxy-4-(trifluoromethoxy)picolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(78.0 mg, 94.0 μmol) and LiCl (16.9 mg, 397 μmol) in DMF (1 mL) washeated in the MW at 200° C. for 10 minutes. The dark brown RM wasdiluted with water and extracted twice with EtOAc and once with 10% MeOHin EtOAc. The combined organic layers were dried over Na₂SO₄,concentrated and dried under reduced pressure. The crude product waspurified by reverse phase preparative HPLC (RP-HPLC basic 1: 5 to 70% Bin 20 min with a plateau at 70% for 1 min) and lyophilized. The productwas dissolved in EtOH/DCM (1:2) and left to stand in an open flask for24 h. The resulting solid was filtered, washed with Et₂O and dried undervacuum at 30° C. to give the title compound.

LC-MS: Rt=1.20 min; MS m/z [M+H]⁺ 771.4/773.4, m/z [M−H]⁻ 769.4/771.4;UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 11.0 (s, br, 1H), 10.35 (s, br, 1H), 8.15(d, J=5.7 Hz, 1H), 8.05 (d, J=8.5 Hz, 1H), 7.96 (m, 1H), 7.71 (dd, J=2.0Hz, 8.4 Hz, 1H), 7.43 (m, 1H), 6.83 (m, 1H), 5.31 (s, 2H), 4.54 (m, 1H),4.25 (m, 2H), 3.80 (m, 2H), 3.48 (m, 3H), 3.25 (m, 1H), 3.00 (m, 3H),2.81 (m, 1H), 2.66 (m, 1H), 2.50 (m, 2H), 1.19 (t, J=7.1 Hz, 3H)

Example 99:N-(2-chloro-5-fluoro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

N-(2-chloro-5-fluoro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate BQ) (390 mg, 580 μmol) in DCM (7 mL) was cooled to 0° C.under argon, then 3-hydroxypicolinoyl chloride (Intermediate CV) (183mg, 1.16 mmol) was added, followed by DIPEA (355 μL, 2.03 mmol). The RMwas stirred at RT for 1.5 hours. The RM was quenched with 5%NaHCO₃-solution, extracted three times with DCM, the organic layer wasdried through a phase separator and concentrated under reduced pressure.The crude product was purified by column chromatography (RediSep Column:Silica 80 g Gold, eluent DCM:MeOH 100:0 to 93:07) to give the titlecompound as a solid.

LC-MS: Rt=1.02 min; MS m/z [M+H]⁺ 706.2/708.3, m/z [M−H]⁻ 704.2/706.2;UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.69 (s, 1H), 10.38 (s, 1H), 8.62 (d,J=11.9 Hz, 1H), 8.06 (m, 1H), 7.29 (m, 2H), 6.81 (m, 1H), 5.40 (s, 2H),4.55 (m, 1H), 4.24 (m, 2H), 3.80 (m, 2H), 3.45 (m, 3H), 3.22 (m, 1H),2.97 (m, 3H), 2.79 (m, 1H), 2.62 (m, 1H), 2.52 (m, 2H), 1.17 (t, J=7.4Hz, 3H)

Example 100:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(pentafluorosulfanyl)phenyl)acetamide

Step 1:2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(pentafluorosulfanyl)phenyl)acetamide

2-(2-(3,6-Dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(pentafluorosulfanyl)phenyl)acetamide(Intermediate BR) (414 mg, 686 μmol),5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (Intermediate CY)(184 mg, 754 μmol) and HATU (417 mg, 1.10 mmol) were mixed in DCM (6 mL)and DIPEA (599 μL, 3.43 mmol) was added. The RM was stirred at RT for 1hour. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL) were added.The aqueous layer was washed twice with DCM (2×10 mL). The combinedorganic layers were dried through a phase separator and concentratedunder reduced pressure. The crude product was purified by columnchromatography (RediSep Column: Silica 24 g, eluent DCM:DCM/MeOH (1/1)100:0 to 72:28) to give the title compound.

LC-MS: Rt=1.22 min; MS m/z [M+H]⁺ 830.5, m/z [M−H]⁻ 828.5; UPLC-MS 1

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(pentafluorosulfanyl)phenyl)acetamide

2-(6-(4-(5-(Benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(pentafluorosulfanyl)phenyl)acetamide(641 mg, 649 μmol) was dissolved in DCM (15 mL) and TFA (1.00 mL, 13.0mmol) was added. The RM was stirred at 45° C. for 1 day. No product wasobserved. TFA (2.00 mL, 26.0 mmol) was added, followed by DCM (2 mL) andthe RM was stirred at 50° C. for 8 hours. Then it was allowed to cool toRT and stirred at RT for 4.5 days. Then TFA (1.00 mL, 13.0 mmol) wasadded and it was stirred at 65° C. for 1 day. The RM was concentratedunder reduced pressure. The crude product was purified in 2 portions byreverse phase preparative HPLC (RP-HPLC acidic 1: 5 to 95% B in 20 minwith a plateau at 95% for 1 min and RP-HPLC acidic 1: 5 to 95% B in 20min with a plateau at 95% for 1 min). The product containing fractionswere basified with aq sat NaHCO₃, extracted twice with DCM, driedthrough a phase separator and concentrated under reduced pressure. Theresidue was suspended in MeOH/ACN (1:1) and sonicated for 1 minute. Thenit was filtered and dried under HV to give a solid. The solid wasdissolved in DCM (5 mL) and EtOH (2 mL) and filtered. The filtratecrystallized over the weekend. The crystals were filtered, washed withEt₂O and dried to give the title compound.

LC-MS: Rt=1.07 min; MS m/z [M+H]⁺ 740.3, m/z [M−H]⁻ 738.3; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.22 (s, 1H), 10.03 (s, 1H), 8.58 (s, 1H),7.75 (m, 3H), 6.82 (m, 1H), 5.25 (s, 2H), 4.52 (m, 1H), 4.26 (m, 2H),3.81 (m, 2H), 3.49 (m, 3H), 3.26 (m, 1H), 2.99 (m, 3H), 2.81 (m, 1H),2.64 (m, 1H), 2.51 (m, 2H), 2.44 (s, 3H), 2.37 (s, 3H), 1.19 (t, J=7.1Hz, 3H)

Example 101:N-(2,4-dichlorophenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To a brown solution ofN-(2,4-dichlorophenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate BS) (258 mg, 436 μmol) in DCM (3 mL) under argon at 0° C.was added 3-hydroxypicolinoyl chloride (Intermediate CV) (103 mg, 654μmol). DIPEA (381 μL, 2.18 mmol) was added dropwise. The resulting brownRM was stirred at 0° C. for 5 hours. 3-Hydroxypicolinoyl chloride(Intermediate CV) (103 mg, 654 μmol) was added to the RM at 0° C., whichwas then stirred for 90.5 hours, allowing the temperature to slowly comeback to RT. The RM was diluted with DCM and washed once with water. Theaqueous layer was extracted once with DCM. The combined organic layerswere dried through a phase separator, concentrated and dried undervacuum. The crude product was purified by column chromatography (RediSepColumn: Silica 12 g, eluent DCM:DCM/MeOH (9/1) 100:0 to 60:40). Theproduct containing fractions were combined, concentrated and dried undervacuum to give a beige solid. MeOH was added to the solid and sonicated.The resulting beige milky suspension was filtered. The cake was washedwith MeOH and dried under vacuum to give the title compound as a whitesolid.

LC-MS: Rt=0.95 min; MS m/z [M+H]⁺ 653.3/655.3/657.3, m/z [M−H]⁻651.5/653.5/655.5; UPLC-MS 6

¹H NMR (600 MHz, DMSO-d₆) δ 10.52 (s, br, 1H), 10.21 (s, br, 1H), 8.05(m, 1H), 7.71 (m, 2H), 7.41 (dd, J=2.4 Hz, 8.9 Hz, 1H), 7.27 (m, 2H),6.83 (m, 1H), 5.24 (s, 2H), 4.53 (m, 1H), 4.26 (m, 2H), 3.80 (m, 2H),3.45 (m, 3H), 3.22 (m, 1H), 2.96 (m, 3H), 2.79 (m, 1H), 2.60 (m, 1H),2.50 (m, 2H), 1.18 (t, J=7.4 Hz, 3H)

Example 102:N-(5-chloro-2-methyl-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

3-Hydroxypicolinic acid (69.3 mg, 488 μmol) was dissolved in DCM (2.5mL) at RT under argon. 1-chloro-N,N,2-trimethylprop-1-en-1-amine (71.7mg, 537 μmol) was added and the RM was stirred at RT for 1.25 hours. Asolution ofN-(5-chloro-2-methyl-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate BT) (175 mg, 244 μmol) in DCM (1.5 mL) and DIPEA (128 μL,732 μmol) was added. The brown solution was stirred at RT for 1.75hours. The RM was quenched with water (4 mL) and aq sat NaHCO₃ (4 mL).Then it was extracted 4 times with DCM (4×40 mL). The organic layer waswashed with water (10 mL), dried through a phase separator andconcentrated under reduced pressure. The crude product was purified byreverse phase preparative HPLC (RP-HPLC acidic 1: 15 to 85% in 20 minwith a plateau at 85% for 1 min). The product containing fractions werecombined and filtered through a PL-HCO₃ MP SPE cartridge (500 mg per 6mL). The filtrate was lyophilized. Repurified by reverse phasepreparative HPLC (RP-HPLC acidic 1: 20 to 80% in 20 min with a plateauat 80% for 1 min). The product containing fractions were combined andfiltered through a PL-HCO₃ MP SPE cartridge (500 mg per 6 mL). Thefiltrate was lyophilized to give the title compound as a colourlesssolid.

LC-MS: Rt=0.99 min; MS m/z [M+H]⁺ 702.4/704.4, m/z [M−H]⁻ 700.4/702.4;UPLC-MS 3

¹H NMR (600 MHz, DMSO-d₆) δ 10.40 (s, br, 2H), 8.50 (s, 1H), 8.06 (m,1H), 7.29 (m, 2H), 6.82 (m, 1H), 5.15 (s, br, 2H), 4.55 (m, 1H), 4.25(m, 2H), 3.81 (m, 2H), 3.49 (m, 2H), 3.42 (m, 1H), 3.22 (m, 1H), 2.96(m, 3H), 2.80 (m, 1H), 2.61 (m, 1H), 2.53 (m, 5H), 1.18 (t, J=7.6 Hz,3H)

Example 103:N-(4-chloro-2-methyl-5-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

N-(4-chloro-2-methyl-5-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate BU) (214 mg, 280 μmol) was dissolved in DCM (2.8 mL) at RTunder argon. 3-Hydroxypicolinoyl chloride (Intermediate CV) (66.3 mg,421 μmol) was added, followed by DIPEA (147 μL, 841 μmol) and the RM wasstirred at RT for 21 hours. 3-Hydroxypicolinoyl chloride (IntermediateCV) (40.0 mg, 254 μmol) and DIPEA (90.0 μL, 515 μmol) were added againand the RM was stirred at RT for 3 hours. The RM was quenched with water(4.00 mL) and aq sat NaHCO₃ (4 mL), then it was extracted with DCM (4×25mL). The organic layer was washed with water (10 mL), dried through aphase separator, concentrated and dried under reduced pressure. Thecrude product was purified by reverse phase preparative HPLC (RP-HPLCacidic 1: 10 to 90% B in 20 min with a plateau at 90% for 1 min). Theproduct containing fractions were combined, basified with aq sat NaHCO₃and the ACN was removed under reduced pressure. The aqueous layer waswashed three times with DCM. The organic layer was dried through a phaseseparator, concentrated and dried under reduced pressure to give a beigesolid. The beige solid was dissolved in MeOH (1.5 mL) and DCM (2 mL) andfiltered. The filtrate crystallized on standing at RT to give the titlecompound as a beige solid.

LC-MS: Rt=1.05 min; MS m/z [M+H]⁺ 701.5/703.5, m/z [M−H]⁻ 699.3/701.4;UPLC-MS 1

¹H NMR (600 MHz, DMSO-d₆) δ 10.38 (s, 1H), 10.07 (s, 1H), 8.06 (m, 1H),7.96 (s, 1H), 7.66 (s, 1H), 7.28 (m, 2H), 6.82 (m, 1H), 5.22 (s, 2H),4.54 (m, 1H), 4.25 (m, 2H), 3.81 (m, 2H), 3.46 (m, 3H), 3.22 (m, 1H),2.98 (m, 3H), 2.80 (m, 1H), 2.63 (m, 1H), 2.50 (m, 2H), 2.34 (s, 3H),1.19 (t, J=7.4 Hz, 3H)

Example 104:N-(5-chloro-4-ethyl-2-methylphenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

N-(5-chloro-4-ethyl-2-methylphenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate BV) (383 mg, 702 μmol) was dissolved in DCM (5 mL). Then3-hydroxypicolinoyl chloride (Intermediate CV) (221 mg, 1.40 mmol) wasadded, followed by DIPEA (490 μL, 2.81 mmol). The RM was stirred at RTfor 1 hour. 3-hydroxypicolinoyl chloride (Intermediate CV) (1.50 g, 9.52mmol) was added, followed by DIPEA (2.45 mL, 14.0 mmol). The RM wasstirred at RT for 2 hours. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM(10 mL) were added. The aqueous layer was washed twice with DCM (2×10mL). The combined organic layers were dried through a phase separatorand concentrated under reduced pressure. The crude product was purifiedby column chromatography (RediSep Column: Silica 40 g, eluentDCM:DCM/MeOH (9/1) 100:0 to 50:50). The product containing fractionswere combined, concentrated under vacuum and dried under HV. The residuewas purified by reverse phase preparative HPLC (RP-HPLC acidic 1: 5 to100% in 20 min). The product containing fractions were combined,basified with aq sat NaHCO₃, extracted twice with DCM, dried through aphase separator and concentrated under reduced pressure to give a whitesolid. The solid was crystallized from DMF (1 mL) to give the titlecompound as a colourless solid.

LC-MS: Rt=1.05 min; MS m/z [M+H]⁺ 661.4/663.4, m/z [M−H]⁻ 659.7/661.7;UPLC-MS 6

¹H NMR (400 MHz, DMSO-d₆) δ 10.39 (s, 1H), 9.84 (s, 1H), 8.07 (m, 1H),7.48 (s, 1H), 7.29 (m, 2H), 7.21 (s, 1H), 6.84 (m, 1H), 5.18 (s, 2H),4.55 (m, 1H), 4.27 (m, 2H), 3.82 (m, 2H), 3.45 (m, 3H), 3.23 (m, 1H),2.99 (m, 3H), 2.80 (m, 1H), 2.62 (m, 3H), 2.55 (m, 2H), 2.22 (s, 3H),1.19 (d, J=7.2 Hz, 3H), 1.14 (t, J=7.4 Hz, 3H)

Example 105:N-(4-chloro-2-methylphenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-5-methyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

3-Hydroxypicolinic acid (205 mg, 1.4 mmol) was dissolved in CM (10 mL)and 1-chloro-N,N,-trimethylprop-1-en-1-amine (260 μL, 1.97 mmol) wasadded. The mixture was stirred at RT for 1.5 hours. ThenN-(4-chloro-2-methylphenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate BW) (550 mg, 983 μmol) and DIPEA (687 μL, 3.93 mmol) wereadded and the mixture was stirred at RT for 20 hours. Water, aq satNaHCO₃ and DCM were added. The aqueous layer was washed twice with DCM.The combined organic layers were dried through a phase separator andconcentrated under reduced pressure. The crude product was purified in 2portions by reverse phase preparative HPLC (RP-HPLC acidic 1: 10 to 90%in 25 min and RP-HPLC acidic 1: 10 to 75% in 22 min). The productcontaining fractions were combined, basified with aq sat NaHCO₃,extracted twice with DCM, dried through a phase separator andconcentrated under reduced pressure to give the title compound.

LC-MS: Rt=0.91 min; MS m/z [M+H]⁺ 619.3/621.3, m/z [M−H]⁻ 617.3/619.3;UPLC-MS 4

¹H NMR (400 MHz, DMSO-do) δ 10.39 (s, 1H), 9.88 (s, 1H), 8.06 (m, 1H),7.43 (d, J=8.5 Hz, 1H), 7.33 (d, J=2.3 Hz, 1H), 7.29 (m, 2H), 7.23 (dd,J=2.4 Hz, 8.7 Hz, 1H), 6.84 (m, 1H), 5.20 (s, 2H), 4.53 (m, 1H), 4.27(m, 2H), 3.81 (m, 2H), 3.43 (m, 3H), 3.23 (m, 1H), 2.98 (m, 1H), 2.77(m, 1H), 2.60 (m, 1H), 2.59 (s, 3H), 2.51 (m, 2H), 2.23 (s, 3H)

Example 106:N-(4-bromo-2-chlorophenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-bromo-2-chlorophenyl)acetamide

N-(4-bromo-2-chlorophenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate BX) (280 mg, 388 μmol) and5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (Intermediate CY)(95.0 mg, 388 μmol) were mixed in DMF (2.1 mL) at RT under argon. HATU(177 mg, 466 μmol) was added, followed by DIPEA (339 μL, 1.94 mmol) andthe RM was stirred at RT for 1.75 hours. The RM was quenched with water(10 mL) and EtOAc (40 mL). Then it was extracted with EtOAc (3×50 mL),water (3×10 mL) and brine (2×10 mL). The organic layer was dried overNa₂SO₄, filtered and concentrated under reduced pressure. The crudeproduct was mixed with EtOH, stirred at 40° C. and sonicated for 1 hour.The suspension was filtered, washed with a small amount of EtOH anddried under HV to give the title compound as a white solid

LC-MS: Rt=1.14 min; MS m/z [M+H]⁺ 802.3/804.3/806.3, m/z [M−H]⁻800.3/802.3/804.3; UPLC-MS 1

Step 2:N-(4-bromo-2-chlorophenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

2-(6-(4-(5-(Benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-bromo-2-chlorophenyl)acetamide(255 mg, 318 μmol) was dissolved in TFA (1.76 mL, 22.9 mmol) and the RMwas stirred at 50° C. for 4.5 hours, then at RT overnight. The RM wasconcentrated under reduced pressure. The resulting foam was extractedwith DCM (4×50 mL), aq sat NaHCO₃ (15 mL) and brine (15 mL). The organiclayer was dried through a phase separator and concentrated under reducedpressure. The crude product was suspended in EtOH (4 mL) and stirred at45° C. for 6 hours, then it was sonicated. DCM was added until fulldissolution and then DCM was carefully removed under reduced pressure.Concentration of the EtOH solution at 45° C. under reduced pressure wascontinued until a cloudiness was observed, then it was stirred at RT for4 hours. The resulting suspension was filtered and dried to give thetitle compound as a beige solid.

LC-MS: Rt=0.98 min; MS m/z [M+H]⁺ 712.2/714.2/716.2, m/z [M−H]⁻710.2/712.2/714.2; UPLC-MS 1

LC-MS: Rt=4.86 min; MS m/z [M+H]⁺ 712.1/714.1/716.1, m/z [M−H]⁻710.1/712.2/714.2; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.22 (s, br, 1H), 10.17 (s, 1H), 8.57 (s,1H), 7.81 (m, 1H), 7.67 (d, J=8.5 Hz, 1H), 7.54 (dd, J=2.1 Hz, 8.8 Hz,1H), 6.83 (m, 1H), 5.24 (s, 2H), 4.52 (m, 1H), 4.26 (m, 2H), 3.80 (m,2H), 3.48 (m, 3H), 3.26 (m, 1H), 2.97 (m, 3H), 2.81 (m, 1H), 2.64 (m,1H), 2.50 (m, 2H), 2.44 (s, 3H), 1.18 (t, J=7.1 Hz, 3H)

Example 107:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide

Step 1:2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide

2-(2-(3,6-Dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide(Intermediate BP) (268 mg, 439 μmol) and5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (Intermediate CY)(107 mg, 439 μmol) were mixed in DMF (2.4 mL) at RT under argon. HATU(200 mg, 527 μmol) was added, followed by DIPEA (383 μL, 2.20 mmol) andthe RM was stirred at RT for 55 minutes. Water (10 mL) and EtOAc (40 mL)was added. Then it was extracted with EtOAc (3×40 mL), water (2×10 mL)and brine (2×10 mL). The organic layer was dried over Na₂SO₄ andconcentrated under reduced pressure. The crude product was mixed withEtOH (4 mL) and stirred at 40° C. The beige suspension was filtered,washed with a small amount of cold EtOH and dried to give the titlecompound as a beige solid.

LC-MS: Rt=1.16 min; MS m/z [M+H]⁺ 776.3, m/z [M−H]⁻ 774.4; UPLC-MS 1

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide

2-(6-(4-(5-(Benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide(199 mg, 257 μmol) was mixed with TFA (1.50 mL, 19.5 mmol) and the RMwas stirred at 50° C. for 20 hours. The RM was concentrated underreduced pressure and dried under HV. The foam was extracted with DCM(4×40 mL), aq sat NaHCO₃ (10 mL) and brine (10 mL). The organic layerwas dried through a phase separator and concentrated under reducedpressure. The crude product was suspended in EtOH (3 mL) and stirred at60° C. for 16 hours. The suspension was filtered and dried under HV togive the title compound as a beige solid.

LC-MS: Rt=1.00 min; MS m/z [M+H]⁺ 686.3, m/z [M−H]⁻ 684.4; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.67 (s, 1H), 10.22 (s, 1H), 8.58 (s, 1H),8.22 (t, J=8.0 Hz, 1H), 7.79 (d, J=11.2 Hz, 1H), 7.56 (d, J=8.3 Hz, 1H),6.80 (m, 1H), 5.30 (s, 2H), 4.52 (m, 1H), 4.24 (m, 2H), 3.79 (m, 2H),3.50 (m, 3H), 3.25 (m, 1H), 2.97 (m, 3H), 2.82 (m, 1H), 2.64 (m, 1H),2.50 (m, 2H), 2.44 (s, 3H), 1.18 (t, J=7.5 Hz, 3H)

Example 108:N-(2-bromo-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-bromo-4-(trifluoromethyl)phenyl)acetamide

N-(2-bromo-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate BY) (280 mg, 358 μmol) and5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (Intermediate CY)(87.0 mg, 358 μmol) were mixed in DMF (2 mL) at RT under argon. HATU(163 mg, 429 μmol) was added, followed by DIPEA (312 μL, 1.79 mmol) andthe RM was stirred at RT for 1.75 hours. Water (10 mL) and EtOAc (40 mL)were added. Then it was extracted with EtOAc (3×40 mL). The combinedorganic layers were washed with water (3×10 mL) and brine (2×10 mL),dried over Na₂SO₄ and concentrated under reduced pressure. The crudeproduct was mixed with EtOH (4.00 mL) and stirred at 40° C. for 5 hours.The suspension was filtered, washed with a small amount of cold EtOH anddried under HV to give the title compound as a white solid.

LC-MS: Rt=1.19 min; MS m/z [M+H]⁺ 836.4/838.5, m/z [M−H]⁻ 834.1/836.0;UPLC-MS 1

Step 2:N-(2-bromo-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

2-(6-(4-(5-(Benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-bromo-4-(trifluoromethyl)phenyl)acetamide(224 mg, 268 μmol) and TFA (1.50 mL, 19.5 mmol) were stirred at 50° C.for 17.5 hours. The RM was concentrated under reduced pressure and driedunder HV. The foam was extracted with DCM (4×40 mL), aq sat NaHCO₃ (10mL) and brine (10 mL). The organic layer was dried through a phaseseparator and concentrated under reduced pressure. The solid wassuspended in EtOH (3 mL) and stirred at 45° C. for 6 hours. Thesuspension was filtered and dried under HV to give the title compound asa beige solid.

LC-MS: Rt=1.03 min; MS m/z [M+H]⁺ 746.2/748.2, m/z [M−H]⁻ 744.3/746.2;UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.26 (s, 1H), 10.22 (s, 1H), 8.58 (s, 1H),8.08 (s, 1H), 7.91 (d, J=8.3 Hz, 1H), 7.76 (d, J=8.3 Hz, 1H), 6.85 (m,1H), 5.29 (s, 2H), 4.52 (m, 1H), 4.26 (m, 2H), 3.80 (m, 2H), 3.49 (m,3H), 3.23 (m, 1H), 2.98 (m, 3H), 2.82 (m, 1H), 2.64 (m, 1H), 2.50 (m,2H), 2.44 (s, 3H), 1.20 (t, J=7.6 Hz, 3H)

Example 109:2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-2-(1,4,5,6-tetrahydropyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Step 1: Tert-butyl5-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)-3,4-dihydropyridine-1(2H)-carboxylate

2-(2-Bromo-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate BZ) (218 mg, 329 μmol), tert-butyl6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydropyridine-1(2H)-carboxylate(Intermediate DI) (152 mg, 493 μmol) and XPhos Pd G3 (13.9 mg, 16.0μmol) were combined in a MW vial. 1,4-Dioxane (3 mL) and 1M aq K₃PO4(986 μL, 986 μmol) were added. The vial was capped, degassed and flushedwith argon. The RM was stirred at 90° C. for 55 minutes. The RM wascooled to RT. The RM was diluted with DCM (8 mL) and water (2 mL). Thebiphasic mixture was stirred at RT for 1 minute then the organic layerwas separated by filtering through a phase separator. AddedSiliaMetS®Thiol and stirred at RT for 15 minutes. Filtered and thefiltrate was concentrated under reduced pressure. The crude product waspurified by reverse phase preparative HPLC (RP-HPLC acidic 1: 40 to 80%in 15 min with a plateau at 80% for 1 min). The product containingfractions were basified with 5% aq NaHCO₃ and extracted with DCM (2×10mL). The organics were eluted through a phase separator and evaporatedin vacuo to give the title compound as a pale yellow gum.

LC-MS: Rt=1.24 min; MS m/z [M+H]⁺ 766.5, m/z [M−H]⁻ 764.7; UPLC-MS 1

Step 2:2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-2-(1,4,5,6-tetrahydropyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To tert-butyl5-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)-3,4-dihydropyridine-1(2H)-carboxylate(60.0 mg, 78.0 μmol) in DCM (2 mL) was added 4N HCl in 1,4-dioxane (3mL) and the suspension was briefly heated with a heat gun to reflux fora few seconds then allowed to stand for 15 minutes. The RM wasconcentrated under reduced pressure The crude product was purified byreverse phase preparative HPLC (RP-HPLC acidic 1: 20 to 55% B in 7 minwith a plateau at 55% for 1 min). The product containing fraction wasbasified with 5% aq NaHCO₃ and extracted with DCM (20 mL). The organiclayer was separated by filtration through a phase separator andevaporated in vacuo to give a yellow solid. It was recrystallized fromEtOAc to give the title compound as a colourless solid.

LC-MS: Rt=0.96 min; MS m/z [M+H]⁺ 666.5, m/z [M−H]⁻ 664.4; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.37 (s, 1H), 9.97 (s, 1H), 8.06 (m, 1H),7.71 (d, J=8.1 Hz, 1H), 7.62 (m, 1H), 7.52 (dd, J=2.3 Hz, 8.5 Hz, 1H),7.29 (m, 3H), 6.11 (m, 1H), 5.16 (s, 2H), 4.54 (m, 1H), 3.45 (m, 3H),3.20 (m, 1H), 3.13 (m, 2H), 2.94 (m, 3H), 2.76 (m, 1H), 2.58 (m, 1H),2.38 (m, 2H), 2.35 (s, 3H), 1.77 (m, 2H), 1.17 (t, J=7.1 Hz, 3H)

Example 110:2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-2-(1,4,5,6-tetrahydropyridin-2-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Step 1: Tert-butyl6-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)-3,4-dihydropyridine-1(2H)-carboxylate

2-(2-Bromo-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate BZ) (2.48 g, 3.74 mmol), tert-butyl6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydropyridine-1(2H)-carboxylate(Intermediate DI) (2.54 g, 8.22 mmol) and XPhos Pd G3 (158 mg, 187 μmol)were mixed in a MW vial. 1,4-Dioxane (30 mL) and 1M aq K₃PO₄ (11.2 mL,11.2 mmol) were added. The vial was capped, degassed and flushed withargon. Then it was stirred at 90° C. for 3 days. The reaction wasallowed to cool to RT. The RM was diluted with DCM (40 mL) and water (10mL). The organic layer was separated by filtering through a phaseseparator. SiliaMetS®Thiol was added, the suspension was stirred at 35°C. for 30 minutes, filtered and the filtrate was concentrated in vacuo.The crude product was purified by column chromatography (RediSep Column:Silica 40 g, eluent DCM:MeOH 100:0 to 95:5) to give the title compoundas a pale brown gum.

LC-MS: Rt=1.14 min; MS m/z [M+H]⁺ 766.5, m/z [M−H]⁻ 764.6; UPLC-MS 1

Step 2:2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-2-(1,4,5,6-tetrahydropyridin-2-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Tert-butyl6-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)-3,4-dihydropyridine-1(2H)-carboxylate(1.20 g, 32% pure) was dissolved in DCM (5 mL) and 4M HCl in 1,4-dioxane(5 mL) was added. Immediately, an insoluble brown gum precipitated.Heated briefly to reflux with a heat gun then allowed to cool for 10minutes. The RM was concentrated in vacuo and the brown residue waspartitioned between DCM (70 mL) and 1M aq HCl (100 mL). The biphasicmixture was stirred vigorously for 10 minutes. The aqueous layer wasseparated, the pH adjusted to pH 5 by addition of solid KOH andextracted with DCM (2×50 mL). The combined DCM layers were elutedthrough a phase separator and concentrated in vacuo to give a pale brownfoam. The crude product was purified by column chromatography (RediSepColumn: Silica 12 g, eluent DCM:MeOH 100:0 to 97:03). The crude productwas further purified by reverse phase preparative HPLC (RP-HPLC acidic1: 5 to 40% in 15 min with a plateau at 40% for 1 min, 25 mL/min). Theproduct containing fractions were basified with aq sat NaHCO₃ andextracted with DCM/MeOH (9/1, 2×30 mL). The combined extracts wereeluted through a phase separator and concentrated under reduced pressureto give a colourless solid. Recrystallized from MeOH/water to give thetitle compound as colourless crystals.

LC-MS: Rt=0.80 min; MS m/z [M+H]⁺ 666.4, m/z [M−H]⁻ 664.5; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.38 (s, 1H), 9.99 (s, 1H), 8.07 (m, 1H),7.76 (m, 1H), 7.63 (m, 1H), 7.53 (m, 1H), 7.29 (m, 2H), 5.45 (m, 1H),5.26 (d, J=23.7 Hz, 2H), 4.83 (m, 1H), 4.56 (m, 1H), 3.75 (m, 1H), 3.46(m, 3H), 3.18 (m, 2H), 2.98 (m, 3H), 2.81 (m, 1H), 2.65 (m, 1H), 2.37(d, J=3.3 Hz, 3H), 2.18 (m, 1H), 1.75 (m, 2H), 1.61 (m, 1H), 1.20 (t,J=7.4 Hz, 3H)

Example 111:rac-2-(5-ethyl-2-(5-fluoro-1,4,5,6-tetrahydropyridin-2-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Step 1: rac-tert-butyl6-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)-3-fluoro-3,4-dihydropyridine-1(2H)-carboxylate

2-(2-Bromo-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate BZ) (91.0 mg, 138 μmol) and XPhos Pd G3 (5.82 mg, 6.88μmol) were combined in a MW vial, tert-butyl3-fluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydropyridine-1(2H)-carboxylate(Intermediate DJ) (45.0 mg, 138 μmol) in 1,4-dioxane (1 mL) and 1M aqK₃PO₄ (413 μL, 413 μmol) were added. The vial was capped, degassed,flushed with argon and stirred at 90° C. for 3 hours. The RM was allowedto cool to RT then diluted with DCM (8 mL) and water (2 mL). Thebiphasic mixture was stirred at RT for 1 minute then the organic layerwas separated by filtering through a phase separator. SiliaMetS®Thiolwas added and the mixture was stirred at RT for 20 minutes. Thesuspension was filtered and the filtrate was concentrated in vacuo. Thecrude product was purified by reverse phase preparative HPLC (RP-HPLCacidic 1: 25 to 60% B in 20 min with a plateau at 60% for 1 min). Theproduct containing fractions were basified with 5% aq NaHCO₃ andextracted with DCM (2×10 mL). The organics were eluted through a phaseseparator and evaporated in vacuo to give the title compound as acolourless gum.

LC-MS: Rt=1.10 min; MS m/z [M+H]⁺ 784.5, m/z [M−H]⁻ 782.6; UPLC-MS 1

Step 2:rac-2-(5-ethyl-2-(5-fluoro-1,4,5,6-tetrahydropyridin-2-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To rac-tert-butyl6-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)-3-fluoro-3,4-dihydropyridine-1(2H)-carboxylatein DCM (300 μL) was added TFA (200 μL) and the solution was brieflyheated with a heat gun, then allowed to stand for 20 minutes. The RM wasevaporated in vacuo, resuspended in 5% aq NaHCO₃ (5 mL) and extractedwith DCM (2×5 mL). The organic layer was separated by filtration througha phase separator and evaporated in vacuo to give the title compound asa pale yellow solid.

LC-MS: Rt=0.94 min; MS m/z [M+H]⁺ 684.5, m/z [M−H]⁻ 682.5; UPLC-MS 1

Example 112:2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-2-(1,2,3,6-tetrahydropyridin-4-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Step 1: tert-butyl4-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate

2-(2-Bromo-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate BZ) (1.42 g, 2.14 mmol), tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate(827 mg, 2.68 mmol) and XPhos Pd G3 (91.0 mg, 107 μmol) were mixed in aMW vial and the RM was purged with vacuum and argon several times. DMF(15 mL) and 1M aq K₃PO₄ (2.14 mL, 2.14 mmol) were added and the RM wasstirred at 80° C. for 2.5 hours. The RM was diluted with DCM and 50% aqbrine. Then it was washed three times with brine and dried through aphase separator. The residue was adsorbed onto Isolute and purified bycolumn chromatography (RediSep Column: Silica 80 g, eluent EtOAc:MeOH100:0 to 75:25). The product containing fractions were combined,concentrated and dried at HV to give the title compound.

LC-MS: Rt=1.23 min; MS m/z [M+H]⁺ 766.5, m/z [M−H]⁻ 764.3; UPLC-MS 1

Step 2:2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-2-(1,2,3,6-tetrahydropyridin-4-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Tert-butyl4-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate(1.52 g, 1.79 mmol) was dissolved in DCM (20 mL) and TFA (2.75 mL, 35.7mmol) was added. The RM was stirred at RT for 2 hours. Aq NaHCO₃ wasadded and the pH was adjusted to pH 10. Then it was extracted 5 timeswith DCM, dried through a phase separator and concentrated under reducedpressure to give the title compound.

LC-MS: Rt=0.71 min; MS m/z [M+H]⁺ 666.6, m/z [M−H]⁻ 664.3; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.01 (s, 1H), 8.05 (m, 1H), 7.71 (d, J=8.5Hz, 1H), 7.62 (m, 1H), 7.53 (dd, J=2.2 Hz, 8.6 Hz, 1H), 7.28 (m, 2H),6.83 (m, 1H), 5.24 (s, 2H), 4.55 (m, 1H), 3.47 (m, 3H), 3.39 (m, 1H),3.22 (m, 2H), 2.98 (m, 3H), 2.91 (m, 2H), 2.79 (m, 1H), 2.61 (m, 1H),2.42 (m, 2H), 2.35 (s, 3H), 1.19 (t, J=7.4 Hz, 3H)

Example 113:2-(2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

2-(2-Bromo-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate BZ) (200 mg, 301 μmol),2-(5,6-dihydro-1,4-dioxin-2-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(77.0 mg, 362 μmol), XPhos Pd G3 (12.8 mg, 15.0 μmol) and 1M aq K₃PO₄(904 μL, 904 μmol) were mixed in 1,4-dioxane (5 mL). The RM was stirredat 80° C. for 30 minutes. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM(10 mL) were added. The aqueous layer was extracted with DCM (2×10 mL)then with EtOAc (4×10 mL). The combined organic layers were elutedthrough a phase separator and concentrated under reduced pressure. Thecrude product was dissolved in DCM and EtOAc and SiliaMetS®Thiol wasadded. The mixture was stirred at 40° C. for 1 hour filtered, and thecake was washed with DCM. The filtrate was concentrated under reducedpressure. The crude product was purified twice by reverse phasepreparative HPLC (RP-HPLC acidic 1: 10 to 90% B in 20 min with a plateauat 90% for 1 min and RP-HPLC acidic 1: 40 to 70% B in 10 min with aplateau at 70% for 1 min). The product containing fractions werecombined, basified with aq sat NaHCO₃, extracted twice with DCM, driedthrough a phase separator and concentrated under reduced pressure togive the title compound as a white solid. The solid was redissolved inDMF and concentrated under reduced pressure to give an oily residuewhich was redissolved in MeOH (1 mL) and left to stand at RT for 2 days.The resulting precipitate was filtered, washed twice with Et₂O and driedunder HV to give the title compound as a colourless solid.

LC-MS: Rt=0.95 min; MS m/z [M+H]⁺ 669.5, m/z [M−H]⁻ 667.5; UPLC-MS 1

¹H NMR (600 MHz, DMSO-d₆) δ 10.38 (s, 1H), 9.98 (s, 1H), 8.06 (m, 1H),7.72 (d, J=8.3 Hz, 1H), 7.63 (m, 1H), 7.53 (dd, J=2.4 Hz, 8.6 Hz, 1H),7.28 (m, 2H), 7.09 (s, 1H), 5.21 (s, 2H), 4.54 (m, 1H), 4.19 (m, 4H),3.47 (m, 2H), 3.40 (m, 1H), 3.21 (m, 1H), 2.96 (m, 3H), 2.79 (m, 1H),2.61 (m, 1H), 2.35 (s, 3H), 1.18 (t, J=7.3 Hz, 3H)

Example 114:2-(2-(5,6-dihydro-2H-pyran-3-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

2-(2-Bromo-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate BZ) (168 mg, 254 μmol), XPhos Pd G3 (10.7 mg, 13.0 μmol),2-(5,6-dihydro-2H-pyran-3-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(64.0 mg, 305 μmol) were suspended in 1,4-dioxane (5 mL) and 1M aq K₃PO₄(761 μL, 761 μmol) was added. The suspension was stirred at 80° C. for 2days. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL) were added.The aqueous layer was extracted with DCM (2×10 mL) and EtOAc (2×10 mL).The combined organic layers were eluted through a phase separator andconcentrated under reduced pressure. The crude product was dissolved inDCM and EtOAc and SiliaMetS®Thiol was added. The mixture was stirred at40° C. for 1 hour, filtered, and the cake was washed with DCM. Thefiltrate was concentrated under reduced pressure. The crude product waspurified by column chromatography (RediSep Column: Silica 24 g, eluentDCM:DCM/MeOH (9/1) 100:0 to 60:40). The product containing fractionswere combined, concentrated under vacuum and dried under HV. The residuewas purified twice by reverse phase preparative HPLC (RP-HPLC acidic 1:30 to 70% B in 20 min with a plateau at 70% for 1 min and RP-HPLC acidic1: 40 to 60% B in 20 min with a plateau at 60% for 1 min). The productcontaining fractions were combined, basified with aq sat NaHCO₃,extracted twice with DCM, dried through a phase separator andconcentrated under reduced pressure to give the title compound as asolid. The solid was dissolved in DCM/MeOH and left to stand at RT for 1day. The resulting precipitate was dried under HV to give the titlecompound as a solid.

LC-MS: Rt=0.99 min; MS m/z [M+H]⁺ 667.4, m/z [M−H]⁻ 665.5; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.37 (s, 1H), 10.00 (s, 1H), 8.06 (m, 1H),7.72 (d, J=8.4 Hz, 1H), 7.63 (m, 1H), 7.53 (dd, J=2.1 Hz, 8.5 Hz, 1H),7.28 (m, 2H), 6.91 (m, 1H), 5.24 (s, 2H), 4.54 (m, 1H), 4.45 (m, 2H),3.76 (m, 2H), 3.45 (m, 3H), 3.22 (m, 1H), 2.97 (m, 3H), 2.79 (m, 1H),2.61 (m, 1H), 2.36 (s, 3H), 2.33 (m, 2H), 1.19 (t, J=7.1 Hz, 3H)

Example 115:2-(2-(1-(2,2-difluoroethyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Step 1: tert-butyl4-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate

2-(2-Bromo-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate BZ) (3.60 g, 4.88 mmol), tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate(1.89 g, 6.10 mmol) and XPhos Pd G3 (207 mg, 244 μmol) were mixed andpurged with argon several times. DMF (40 mL) was added, followed byK₃PO₄ 1M in water (4.88 mL, 4.88 mmol). The RM was stirred at 80° C. for4.5 hours. XPhos Pd G3 (207 mg, 244 μmol) was added again and the RM wasstirred at 80° C. for 1.5 hours. K₃PO₄ 1M in water (14.7 mL, 14.7 mmol)was added and the RM was stirred at 80° C. overnight. The RM was dilutedwith DCM and 50% brine. Then it was washed three times with brine anddried through a phase separator. The residue was adsorbed onto Isoluteand purified by column chromatography (RediSep Column: Silica 220 g,eluent EtOAc:MeOH 100:0 to 95:5) to give the title compound as a solid.

LC-MS: Rt=1.19 min; MS m/z [M+H]⁺ 766.5, m/z [M−H]⁻ 764.3; UPLC-MS 1

Step 2:2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-2-(1,2,3,6-tetrahydropyridin-4-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Tert-butyl4-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate(3.27 g, 3.71 mmol) was dissolved in DCM (40 mL) and TFA (5.72 mL, 74.3mmol) was added. The RM was stirred at RT for 3 hours. The RM wasquenched with aq NaHCO₃ and the pH adjusted to pH 10. Then it wasextracted 5 times with DCM, dried through a phase separator andconcentrated under reduced pressure. The crude product was purified byreverse phase preparative ISCO (RediSep Column: C18 240 g, eluentwater:ACN 100:0 to 0:100). The product containing fractions werecombined and lyophilized to give the title compound as a powder.

LC-MS: Rt=0.75 min; MS m/z [M+H]⁺ 666.3, m/z [M−H]⁻ 664.3; UPLC-MS 1

Step 3:2-(2-(1-(2,2-difluoroethyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

DIPEA (803 μL, 4.60 mmol) and 2,2-difluoroethan-1-ol (234 μL, 3.70 mmol)were dissolved in dry DMF (8 mL) and heated up to 40° C. Sulfuryldifluoride was bubbled through the RM for 2 mins, then the RM was purgedwith argon for 10 minutes. This RM was added dropwise to a solution of2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-2-(1,2,3,6-tetrahydropyridin-4-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(600 mg, 901 μmol) in DMF (8 mL) at 40° C. The RM was stirred at 40° C.for 2 hours. The residue was adsorbed onto Isolute and purified byreverse phase preparative ISCO (RediSep Column: C18 100 g Gold, eluentwater:ACN 100:0 to 0:100). The product containing fractions werecombined and concentrated under reduced pressure. The solid was furtherpurified by reverse phase preparative ISCO (RediSep Column: C18 50 gGold, eluent water:ACN 100:0 to 0:100) and lyophilized. The resultingsolid was dissolved in EtOAc:EtOH (1:1) and stirred at 57° C. for 48hours, then at RT over the weekend. Afterwards it was cooled in an icebath and the solid was filtered off and dried under HV to give the titlecompound.

LC-MS: Rt=0.95 min; MS m/z [M+H]⁺ 730.3, m/z [M−H]⁻ 728.3; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.37 (s, 1H), 9.99 (s, 1H), 8.06 (m, 1H),7.71 (d, J=8.6 Hz, 1H), 7.62 (m, 1H), 7.53 (d, J=9.0 Hz, 1H), 7.28 (m,2H), 6.75 (m, 1H), 6.35-6.00 (t, J=55.4 Hz, 1H), 5.23 (s, 1H), 4.54 (m,1H), 3.44 (m, 4H), 3.22 (m, 2H), 3.05-2.75 (m, 11H), 2.61 (m, 1H), 2.35(s, 3H), 1.19 (t, J=7.3 Hz, 3H)

Example 116:N-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

N-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate CA) (212 mg, 228 μmol) was dissolved in DCM (2 mL) and3-hydroxypicolinoyl chloride (Intermediate CV) (64.0 mg, 406 μmol) wasadded, followed by DIPEA (120 μL, 685 μmol). The RM was stirred at RTfor 15 minutes. Then DIPEA (200 μL, 1.15 mmol) was added and the RM wasstirred at RT for 7 hours. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM(10 mL) were added. The aqueous layer was washed twice with DCM (2×10mL). The combined organic layers were dried through a phase separatorand concentrated under reduced pressure. The crude product was purifiedby column chromatography (RediSep Column: Silica 24 g, eluentDCM:DCM/MeOH (9/1) 100:0 to 60:40) to afford the title compound as awhite solid.

LC-MS: Rt=1.08 min; MS m/z [M+H]⁺ 705.4/707.4, m/z [M−H]⁻ 703.5/705.4;UPLC-MS 1

¹H NMR (600 MHz, DMSO-d₆) δ 10.47 (s, 1H), 10.38 (s, 1H), 8.10 (d,J=12.6 Hz, 1H), 8.06 (dd, J=2.2 Hz, 5.8 Hz, 1H), 8.02 (d, J=7.1 Hz, 1H),7.29 (m, 2H), 6.81 (m, 1H), 5.36 (s, 2H), 4.55 (m, 1H), 4.25 (m, 2H),3.80 (m, 2H), 3.45 (m, 3H), 3.22 (m, 1H), 2.97 (m, 3H), 2.80 (m, 1H),2.61 (m, 1H), 2.50 (m, 2H), 1.17 (t, J=7.5 Hz, 3H)

Example 117:2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-5-methyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

3-Hydroxypicolinic acid (53.8 mg, 386 μmol) was mixed with DCM (4 mL) atRT. The mixture was cooled to 0° C. and1-chloro-N,N,2-trimethylprop-1-en-1-amine (56.0 μL, 425 μmol) was addeddropwise. The RM was stirred at RT for 2 hours, then it was cooled to 0°C. again. DIPEA (169 μL, 966 μmol) was added. This solution was addeddropwise at 0° C. to a solution of2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide(Intermediate BN) (100 mg, 193 μmol) in DCM (4 mL). The RM was stirredat RT for 1 hour. The RM was diluted with DCM, washed with aq satNaHCO₃, dried over Na₂SO₄, concentrated and dried. The crude product waspurified by reverse phase preparative HPLC (RP-HPLC acidic 1: 5 to 100%B in 20 min). The product containing fractions were combined and the ACNwas removed under reduced pressure. The aqueous residue was mixed withDCM, basified with NaHCO₃, dried over Na₂SO₄ and concentrated underreduced pressure to give the title compound as a solid.

LC-MS: Rt=0.96 min; MS m/z [M+H]⁺ 639.2, m/z [M−H]⁻ 637.1; UPLC-MS 8

¹H NMR (400 MHz, DMSO-d₆) δ 10.85 (s, 1H), 10.37 (s, 1H), 8.07 (m, 1H),7.78 (d, J=8.6 Hz, 2H), 7.70 (d, J=8.6 Hz, 2H), 7.29 (m, 2H), 6.81 (m,1H), 5.22 (s, 2H), 4.53 (m, 1H), 4.24 (m, 2H), 3.80 (m, 2H), 3.44 (m,3H), 3.22 (m, 1H), 2.98 (m, 1H), 2.79 (m, 1H), 2.62 (m, 1H), 2.57 (s,3H), 2.49 (m, 2H)

Example 118:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((2S,5R)-4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,5-dimethylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-((2S,5R)-4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,5-dimethylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

To a brown solution ofN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate CC) (1.11 g, 1.28 mmol) and5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (Intermediate CY)(369 mg, 1.51 mmol) in DMF (14.4 mL) were added HATU (656 mg, 1.73 mmol)and DIPEA (754 μL, 4.32 mmol) at RT and the RM was stirred at RT for 45minutes. The RM was quenched with water and extracted twice with EtOAc.The combined organic layers were washed three times with brine thendried over MgSO₄ cartridge and concentrated under vacuum. The crudeproduct was purified by column chromatography (Silica gel column: Silica40 g, eluent DCM:DCM/MeOH (8/2) 100:0 to 55:45) to give the titlecompound.

LC-MS: Rt=1.23 min; MS m/z [M+H]⁺ 820.5/822.5, m/z [M−H]⁻ 818.5/820.5;UPLC-MS 1

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((2S,5R)-4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,5-dimethylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

2-(6-((2S,5R)-4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,5-dimethylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(1.09 g, 998 μmol) was dissolved in DCM (3 mL). TFA (3.08 mL, 39.9 mmol)was added. The brown solution was stirred at 45° C. for 18 hours. The RMwas diluted with DCM, washed with water and aq sat NaHCO₃. The aqueouslayer was extracted twice with DCM. The combined organic layers weredried through a phase separator and concentrated under reduced pressure.The crude product was adsorbed onto Isolute and purified by columnchromatography (Silica gel column: Silica 40 g, eluent DCM:DCM/MeOH(8/2) 100:0 to 65:35). The product containing fractions were combinedand concentrated under reduced pressure to give a light brown solid. Thelight brown solid was purified in 2 portions by reverse phasepreparative HPLC (2×RP-HPLC acidic 14: 20 to 70% B in 20 min with aplateau at 70% for 3 min). The product containing fractions werecombined, the ACN was removed and the residue was quenched with aq satNaHCO₃, then extracted twice with DCM. The combined organic layers weredried through a phase separator and concentrated under reduced pressureto give the title compound as a beige solid.

LC-MS: Rt=1.11 min; MS m/z [M+H]⁺ 730.2/732.2, m/z [M−H]⁻ 728.3/730.3;UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.35 (s, 1H), 10.14 (s, br, 1H), 8.57 (m,1H), 8.05 (m, 1H), 7.96 (m, 1H), 7.71 (m, 1H), 6.82 (m, 1H), 5.31 (m,2H), 4.21 (m, 3H), 3.79 (m, 2H), 3.69 (m, 1H), 3.45 (m, 1H), 3.11 (m,2H), 2.87 (m, 1H), 2.45 (m, 5H), 2.33 (m, 1H), 1.35 (m, 2H), 1.24 (m,2H), 1.16 (m, 4H), 1.03 (m, 1H), 0.92 (m, 1H)

Example 119:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((2R,3R)-4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-((2R,3R)-4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

To a stirred solution ofN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-((2R,3R)-2,3-dimethylpiperazin-1-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate CI) (618 mg, 978 μmol),5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (Intermediate CY)(239 mg, 978 μmol) and HATU (446 mg, 1.17 mmol) in DMF (2 mL) was addedDIPEA (854 μL, 4.89 mmol) at RT and the RM was stirred at RT for 10minutes. The RM was diluted with EtOAc/water, extracted twice with EtOAcand the combined organic extracts were washed with brine, dried overNa₂SO₄ and concentrated. The crude product was purified by columnchromatography (RediSep Column: Silica 40 g, eluent DCM:DCM/MeOH (8/2)100:0 to 50:50) to give the title compound as a beige foam.

LC-MS: Rt=1.24 min; MS m/z [M+H]⁺ 820.5/822.5, m/z [M−H]⁻ 818.4/820.5;UPLC-MS 1

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((2R,3R)-4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

2-(6-((2R,3R)-4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(858 mg, 889 μmol) in TFA (5.00 mL, 64.9 mmol) was stirred at 40° C. for14 hours. The RM was diluted with DCM and NaHCO₃, extracted twice withDCM and the combined organic extracts were washed with brine, dried overNa₂SO₄ and concentrated. The crude product was purified by columnchromatography (RediSep Column: Silica 40 g, eluent DCM:DCM/MeOH (8/2)100:0 to 50:50) to give the title compound as a white solid.

LC-MS: Rt=1.12 min; MS m/z [M+H]⁺ 730.1/732.1, m/z [M−H]⁻ 728.3/730.2;UPLC-MS 1

LC-MS: Rt=5.50 min; MS m/z [M+H]⁺ 730.1/732.1, m/z [M−H]⁻ 728.3/730.2;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.36 (s, br, 1H), 10.17 (s, br, 1H), 8.56(m, 1H), 8.06 (m, 1H), 7.96 (m, 1H), 7.71 (m, 1H), 6.82 (m, 1H), 5.34(m, 2H), 4.60-4.35 (m, 1H), 4.25 (m, 2H), 4.09 (m, 1H), 3.80 (m, 2H),3.38 (m, 1H), 3.13 (m, 3H), 2.88 (m, 2H), 2.58 (m, 1H), 2.51 (m, 2H),2.44 (m, 3H), 1.48 (m, 2H), 1.22 (m, 5H), 1.08 (m, 1H)

Example 120:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((2S,3S)-4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-((2S,3S)-4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

To a stirred solution ofN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-((2S,3S)-2,3-dimethylpiperazin-1-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate CJ) (50.0 mg, 84.0 μmol),5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (Intermediate CY)(20.6 mg, 84.0 μmol) and HATU (38.4 mg, 101 μmol) in DMF (1 mL) wasadded DIPEA (74.0 μL, 421 μmol) at RT and the RM was stirred at RT for10 minutes. The RM was diluted with EtOAc/water, extracted twice withEtOAc and the combined organic extracts were washed with brine, driedover Na₂SO₄ and concentrated. The crude product was purified by columnchromatography (RediSep Column: Silica 12 g, eluent DCM:DCM/MeOH (8/2)100:0 to 50:50) to give the title compound as a white foam.

LC-MS: Rt=1.24 min; MS m/z [M+H]⁺ 820.3/822.3, m/z [M−H]⁻ 818.4/820.4;UPLC-MS 1

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((2S,3S)-4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

2-(6-((2S,3S)-4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(57.0 mg, 63.0 μmol) in TFA (1.00 mL, 13.0 mmol) was stirred at 50° C.for 14 hours. The RM was diluted with DCM/water, extracted twice withDCM and the combined organic extracts were washed with brine, dried overNa₂SO₄ and concentrated. The crude product was purified by reverse phasepreparative HPLC (RP-HPLC acidic 1: 5 to 95% B in 20 min with a plateauat 95% for 1 min). The product containing fractions were combined,basified with NaHCO₃, the ACN was removed and the resulting aqueousresidue was extracted with DCM. The combined organic layers were washedwith brine, dried over Na₂SO₄ and concentrated under reduced pressure togive the title compound as a white solid.

LC-MS: Rt=1.11 min; MS m/z [M+H]⁺ 730.3/732.3, m/z [M−H]⁻ 728.4/730.4;UPLC-MS 1

LC-MS: Rt=5.51 min; MS m/z [M+H]⁺ 730.2/732.2, m/z [M−H]⁻ 728.3/730.3;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.37 (s, br, 1H), 10.17 (s, br, 1H), 8.56(m, 1H), 8.06 (m, 1H), 7.97 (m, 1H), 7.71 (m, 1H), 6.82 (m, 1H), 5.35(m, 2H), 4.60-4.35 (m, 1H), 4.25 (m, 2H), 4.10 (m, 1H), 3.80 (m, 2H),3.37 (m, 1H), 3.25-2.80 (m, 4H), 2.57 (m, 1H), 2.51 (m, 2H), 2.44 (m,3H), 2.40 (m, 1H), 1.48 (m, 2H), 1.23 (m, 5H), 1.08 (m, 1H)

Example 121:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((2R,3S)-4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideorN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((2S,3R)-4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-((2R,3S)-4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamideor2-(6-((2S,3R)-4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

To a stirred solution ofN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-((2R,3S)-2,3-dimethylpiperazin-1-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideorN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-((2S,3R)-2,3-dimethylpiperazin-1-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate CK) (212 mg, 353 μmol),5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (Intermediate CY)(86.0 mg, 353 μmol) and HATU (161 mg, 424 μmol) in DMF (2 mL) was addedDIPEA (309 μL, 1.77 mmol) at RT and the RM was stirred at RT for 5minutes. The RM was diluted with EtOAc/water, extracted twice with EtOAcand the combined organic extracts were washed with brine, dried overNa₂SO₄ and concentrated. The crude product was purified by columnchromatography (RediSep Column: Silica 24 g, eluent DCM:DCM/MeOH (8/2)100:0 to 40:60) to give the title compound as a white foam.

LC-MS: Rt=1.25 min; MS m/z [M+H]⁺ 820.4/822.4, m/z [M−H]⁻ 818.4/820.4;UPLC-MS 1

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((2R,3S)-4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideorN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((2S,3R)-4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

2-(6-((2R,3S)-4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamideor2-(6-((2S,3R)-4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(285 mg, 264 μmol) in TFA (1.00 mL, 13.0 mmol) was stirred at 40° C. for20 hours. The RM was diluted with DCM and NaHCO₃, extracted twice withDCM and the combined organic extracts were washed with brine, dried overNa₂SO₄ and concentrated. The crude product was purified by columnchromatography (RediSep Column: Silica 24 g, eluent DCM:DCM/MeOH (8/2)100:0 to 50:50). The product containing fractions were combined andconcentrated to give a yellow foam. The foam was triturated in ACN and awhite solid was filtered. The solid was stirred overnight in 10% waterin EtOH at 40° C. to afford the title compound as a white solid.

LC-MS: Rt=1.09 min; MS m/z [M+H]⁺ 730.2/732.1, m/z [M−H]⁻ 728.3/730.3;UPLC-MS 1

LC-MS: Rt=5.38 min; MS m/z [M+H]⁺ 730.2/732.1, m/z [M−H]⁻ 728.3/730.3;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.38 (s, br, 2H), 8.56 (d, J=5.4 Hz, 1H),8.05 (d, J=8.4 Hz, 1H), 7.96 (d, J=2.1 Hz, 1H), 7.71 (dd, J=1.8 Hz, 8.7Hz, 1H), 6.83 (m, 1H), 5.32 (m, 2H), 4.60-4.35 (m, 1H), 4.25 (m, 2H),3.79 (m, 2H), 3.49 (m, 1H), 3.21 (m, 3H), 2.89 (m, 2H), 2.63 (m, 1H),2.52 (m, 2H), 2.44 (m, 3H), 1.22 (m, 6H), 0.85-0.65 (m, 3H)

Example 122:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((2S,3R)-4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideorN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((2R,3S)-4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-((2S,3R)-4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamideor2-(6-((2R,3S)-4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

To a stirred solution ofN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-((2S,3R)-2,3-dimethylpiperazin-1-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideorN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-((2R,3S)-2,3-dimethylpiperazin-1-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate CL) (212 mg, 343 μmol),5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (Intermediate CY)(84.0 mg, 343 μmol) and HATU (156 mg, 411 μmol) in DMF (2 mL) was addedDIPEA (299 μL, 1.71 mmol) at RT and the RM was stirred at RT for 5minutes. The RM was diluted with EtOAc/water, extracted twice with EtOAcand the combined organic extracts were washed with brine, dried overNa₂SO₄ and concentrated. The crude product was purified by columnchromatography (RediSep Column: Silica 24 g, eluent DCM:DCM/MeOH (8/2)100:0 to 40:60) to give a beige foam.

LC-MS: Rt=1.25 min; MS m/z [M+H]⁺ 820.4/822.3, m/z [M−H]⁻ 818.4/820.4;UPLC-MS 1

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((2S,3R)-4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideorN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((2R,3S)-4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

2-(6-((2S,3R)-4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamideor2-(6-((2R,3S)-4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,3-dimethylpiperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(273 mg, 233 μmol) in TFA (1.00 mL, 13.0 mmol) was stirred at 40° C. for14 hours. The RM was diluted with DCM and NaHCO₃, extracted twice withDCM and the combined organic extracts were washed with brine, dried overNa₂SO₄ and concentrated. The crude product was purified in 2 portions byreverse phase preparative HPLC (RP-HPLC acidic 1: 5 to 80% B in 25 minwith a plateau at 80% for 1 min and RP-HPLC acidic 1: 30 to 70% B in 20min with a plateau at 70% for 1 min). The product containing fractionswere basified with NaHCO₃, the ACN was evaporated and the resultingaqueous residue was extracted with DCM, the organic extract was washedwith brine, dried over Na₂SO₄ and concentrated to afford the titlecompound as a white solid.

LC-MS: Rt=1.10 min; MS m/z [M+H]⁺ 730.3/732.3, m/z [M−H]⁻ 728.4/730.4;UPLC-MS 1

LC-MS: Rt=5.47 min; MS m/z [M+H]⁺ 730.3/732.3, m/z [M−H]⁻ 728.3/730.3;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.37 (s, br, 1H), 10.20 (s, br, 1H), 8.55(d, J=6.1 Hz, 1H), 8.04 (d, J=8.5 Hz, 1H), 7.96 (d, J=2.3 Hz, 1H), 7.71(dd, J=1.9 Hz, 9.0 Hz, 1H), 6.83 (m, 1H), 5.32 (m, 2H), 4.65-4.30 (m,1H), 4.25 (m, 2H), 3.79 (m, 2H), 3.46 (m, 2H), 3.20 (m, 2H), 2.89 (m,2H), 2.63 (m, 1H), 2.51 (m, 2H), 2.44 (m, 3H), 1.22 (m, 6H), 0.85-0.65(m, 3H)

Example 123:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Under nitrogen was mixed 5-hydroxy-6-methylpyrimidine-4-carboxylic acid(Intermediate DB) (2.61 g, 17.0 mmol) in ACN (40 mL) as a suspension. Asuspension of HOAt (192 mg, 1.41 mmol) in ACN was added, followed byEDCI (4.06 g, 21.2 mmol). The mixture was stirred at RT for 1 hour.N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AK) (8.00 g, 14.1 mmol) and DIPEA (7.41 mL, 42.2 mmol)were added. The RM was stirred at RT for 1 hour, then it was treatedwith water (8 mL) and stirred for additional 8 hours. Diluted with THE(160 mL) and 20% NaH₂PO₄ solution (80 mL), the organic phase wasisolated, and washed with brine. The organic layer was further dilutedwith THF (160 mL) and was dried over MgSO₄. The solid was removed byfiltration and was concentrated under vacuum (to remove 25% of solvent).A solution of NaOtBu (1.49 g, 15.6 mmol) in THE was added dropwise, andthe resulting suspension was filtered to give the title compound as asodium salt.

LC-MS: Rt=1.05 min; MS m/z [M+H]⁺ 702.4/704.4, m/z [M−H]⁻ 700.5/702.5;UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.36 (s, br, 2H), 8.55 (s, 1H), 8.05 (d,J=8.5 Hz, 1H), 7.96 (m, 1H), 7.71 (dd, J=2.1 Hz, 8.8 Hz, 1H), 6.83 (m,1H), 5.32 (s, 2H), 4.52 (m, 1H), 4.25 (m, 2H), 3.80 (m, 2H), 3.48 (m,3H), 3.25 (m, 1H), 2.99 (m, 3H), 2.81 (m, 1H), 2.64 (m, 1H), 2.52 (m,2H), 2.43 (s, 3H), 1.18 (t, J=7.4 Hz, 3H)

Example 124:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((1R,6R)-5-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-((1R,6R)-5-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide2-(6-((1R,6R)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(Intermediate CP)

(159 mg, 215 μmol) and 5-(benzyloxy)-6-methylpyrimidine-4-carboxylicacid (Intermediate CY) (55.1 mg, 226 μmol) were mixed in DMF (3 mL) atRT under argon. HATU (101 mg, 258 μmol) was added, followed by DIPEA(188 μL, 1.07 mmol). The RM was stirred at RT for 1.2 hours. The RM wasquenched with water (5 mL), then extracted with EtOAc (3×60 mL), water(2×20 mL) and brine (2×20 mL). The combined organic layers were driedthrough a phase separator and concentrated under reduced pressure togive a brown oil. The crude product was adsorbed onto Isolute andpurified by column chromatography (RediSep Column: Silica 40 g, eluentDCM:DCM/MeOH (8/2) 100:0 to 60:40) to give a beige solid. The impurefractions were concentrated under reduced pressure then adsorbed ontoIsolute and purified by column chromatography (RediSep Column: Silica 24g, eluent DCM:DCM/MeOH (8/2) 100:0 to 60:40). The product containingfractions were combined were combined with the solid obtained from thefirst purification to give the title compound as a beige solid.

LC-MS: Rt=1.22 min; MS m/z [M+H]⁺ 818.5/820.5, m/z [M−H]⁻ 816.5/818.5;UPLC-MS 1

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((1R,6R)-5-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

2-(6-((1R,6R)-5-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(81.0 mg, 94.0 μmol) was mixed with TFA (2.17 mL, 28.2 mmol) and the RMwas stirred at 50° C. for 2.5 hours, then it was concentrated underreduced pressure. The residue was dissolved in DCM and concentratedunder reduced pressure. This was performed twice. The crude product waspurified by reverse phase preparative HPLC (RP-HPLC acidic 1: 25 to 90%B in 20 min with a plateau at 90% for 1 min). The product containingfractions were combined, basified with aq sat NaHCO₃ (3 mL), the ACN wasremoved under reduced pressure and the residue was extracted with DCM(4×50 mL), dried through a phase separator and concentrated underreduced pressure to give the title compound as a beige solid.

Chiral HPLC (C-HPLC 11): Rt=2.65 min, 95% ee

LC-MS: Rt=1.13 min; MS m/z [M+H]⁺ 728.5/730.5, m/z [M−H]⁻ 726.4/728.4;UPLC-MS 1

LC-MS: Rt=5.52 min; MS m/z [M+H]⁺ 728.2/730.3, m/z [M−H]⁻ 726.3/728.3;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.34 (m, 2H), 8.56 (s, 1H), 8.05 (d, J=8.6Hz, 1H), 7.96 (d, J=1.9 Hz, 1H), 7.71 (dd, J=2.1 Hz, 8.5 Hz, 1H), 6.83(m, 1H), 5.31 (s, 2H), 4.41 (m, 1H), 4.25 (m, 2H), 3.80 (m, 2H), 3.62(m, 1H), 3.40 (m, 3H), 3.25 (m, 1H), 3.01 (m, 2H), 2.51 (m, 2H), 2.43(m, 4H), 1.63 (m, 1H), 1.34 (m, 2H), 1.19 (t, J=7.2 Hz, 3H)

Example 125N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((1S,6S)-5-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-((1S,6S)-5-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide2-(6-((1S,6S)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(Intermediate CQ)

(143 mg, 184 μmol) and 5-(benzyloxy)-6-methylpyrimidine-4-carboxylicacid (Intermediate CY) (47.1 mg, 193 μmol) were mixed in DMF (2.5 mL) atRT under argon. HATU (86.0 mg, 220 μmol) was added, followed by DIPEA(160 μL, 918 μmol). The RM was stirred at RT for 45 minutes. The RM wasquenched with water (5 mL), then extracted with EtOAc (3×60 mL), water(2×20 mL) and brine (2×15 mL). The combined organic layers were driedthrough a phase separator and concentrated under reduced pressure togive a bright brown solid. The crude product was adsorbed onto Isoluteand purified by column chromatography (RediSep Column: Silica 40 g,eluent DCM:DCM/MeOH (8/2) 100:0 to 60:40) to give the title compound asa pale beige solid.

LC-MS: Rt=1.23 min; MS m/z [M+H]⁺ 818.5/820.5, m/z [M−H]⁻ 816.5/818.5;UPLC-MS 1

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((1S,6S)-5-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

2-(6-((1S,6S)-5-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(100 mg, 122 μmol) was mixed with TFA (2.83 mL, 36.7 mmol) and the RMwas stirred at 50° C. for 3 hours, then it was concentrated underreduced pressure. The crude product was purified by reverse phasepreparative HPLC (RP-HPLC acidic 1: 20 to 90% B in 20 min with a plateauat 90% for 1 min). The product containing fractions were combined,basified with aq sat NaHCO₃ (3 mL), the ACN was removed under reducedpressure and the residue was extracted with DCM (4×40 mL), dried througha phase separator and concentrated under reduced pressure to give thetitle compound as a white solid. The product was dissolved in EtOAc andleft to stand in a closed flask for 18 hours. The resulting solid wasfiltered, washed with EtOAc and dried at 60° C. under reduced pressureto give a colorless solid. The crystals were combined with the motherliquors and stirred in EtOAc at 50° C. for 18 hours. Filtered and driedat 60° C. under reduced pressure to give a colorless solid.

Chiral HPLC (C-HPLC 12): Rt=3.49 min, 95% ee

LC-MS: Rt=1.12 min; MS m/z [M+H]⁺ 728.3/730.3, m/z [M−H]⁻ 726.4/728.3;UPLC-MS 1

LC-MS: Rt=5.54 min; MS m/z [M+H]⁺ 728.3/730.2, m/z [M−H]⁻ 726.3/728.2;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.34 (m, 2H), 8.56 (s, 1H), 8.05 (d, J=8.7Hz, 1H), 7.96 (d, J=2.0 Hz, 1H), 7.71 (dd, J=2.3 Hz, 8.7 Hz, 1H), 6.83(m, 1H), 5.31 (s, 2H), 4.41 (m, 1H), 4.25 (m, 2H), 3.80 (m, 2H), 3.64(m, 1H), 3.40 (m, 3H), 3.25 (m, 1H), 3.01 (m, 2H), 2.51 (m, 2H), 2.43(m, 4H), 1.63 (m, 1H), 1.34 (m, 2H), 1.19 (t, J=7.1 Hz, 3H)

Example 126: tert-butyl4-(6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-5-methyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate

3-Hydroxypicolinic acid (166 mg, 1.17 mmol) was dissolved in DCM (9.5mL) at RT under argon. 1-Chloro-N,N,2-trimethylprop-1-en-1-amine (170μL, 1.29 mmol) was added and the RM was stirred at RT for 1.5 hours.Tert-butyl4-(5-methyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-6-(piperazin-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate(Intermediate CR) (380 mg, 585 μmol) was added, followed by DIPEA (511μL, 2.93 mmol) and the RM was stirred at RT for 4 hours and quenchedwith water. Aq sat NaHCO₃ (10 mL) was added and the mixture wasextracted with DCM (4×30 mL). The combined organic layers were driedthrough a phase separator and concentrated under reduced pressure togive a green-brown oil. The oil was adsorbed onto Isolute and purifiedby column chromatography (RediSep Column: Silica 24 g, eluent DCM:MeOH100:0 to 70:30) to give the title compound as a beige solid.

LC-MS: Rt=1.09 min; MS m/z [M+H]⁺ 738.3, m/z [M−H]⁻ 736.1; UPLC-MS 4

Example 127:(S)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoro-3,4-dihydro-2H-pyran-6-yl)-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:(S)-2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-5-ethyl-2-(3-fluoro-3,4-dihydro-2H-pyran-6-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

To a stirred solution of 5-(benzyloxy)-6-methylpyrimidine-4-carboxylicacid (Intermediate CY) (5.86 mg, 24.0 μmol) and HATU (10.0 mg, 26.0μmol) in DMF (1 mL) was added DIPEA (21.0 μL, 120 μmol) at RT and the RMwas stirred at RT for 10 minutes. Then a solution of(S)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoro-3,4-dihydro-2H-pyran-6-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate CS) (14.0 mg, 24.0 μmol) in DMF (1 mL) was added dropwiseat RT. The RM was stirred at RT for 1 hour. The RM was extracted withwater (30 mL) and TBME (30 mL). The organic phase was washed with aq satNaHCO₃ (30 mL) and brine (30 mL). The combined aqueous layers wereextracted with TBME (2×30 mL). The combined organic phases were driedover anhydrous Na₂SO₄, filtered and concentrated in vacuo at 45° C. togive a yellow resin. The crude product was purified by columnchromatography (Silica gel column: Silica 24 g SNAP cartridge (Biotage),eluent heptane:EtOAc 80:20 to 0:100, then DCM:DCM/MeOH (9/1) 100:0 to0:100) to give the title compound as a yellow resin.

LC-MS: Rt=1.19 min; MS m/z [M+H]⁺ 810.4/812.4, m/z [M−H]⁻ 808.4/810.4;UPLC-MS 1

Step 2:(S)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoro-3,4-dihydro-2H-pyran-6-yl)-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

(S)-2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-5-ethyl-2-(3-fluoro-3,4-dihydro-2H-pyran-6-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(11.4 mg, 11.0 μmol) in TFA (1 mL) was heated to 50° C. and stirred for4 hours. The RM was diluted with DCM (5 mL) and evaporated under reducedpressure at 45° C. The residue was dissolved in DCM (3×5 mL) andevaporated again to give the crude product as TFA salt. This crudeproduct was purified by reverse phase preparative HPLC (RP-HPLC basic 1:5 to 100% B in 20 min). The product containing fractions were combinedand concentrated under reduced pressure to give a yellow solid. Theyellow solid was purified by reverse phase preparative HPLC (RP-HPLCbasic 1: 5 to 70% B in 20 min with a plateau at 70% for 1 min) to givethe title compound as a yellow resin.

LC-MS: Rt=1.03 min; MS m/z [M+H]⁺ 720.4/722.4, m/z [M−H]⁻ 718.3/720.3;UPLC-MS 1

¹H NMR (600 MHz, DMSO-d₆) δ 10.36 (s, br, 1H), 8.73 (m, br, 1H), 8.47(s, 1H), 8.09 (d, J=9.0 Hz, 1H), 7.97 (d, J=1.9 Hz, 1H), 7.71 (dd, J=2.1Hz, 8.7 Hz, 1H), 5.83 (m, 1H), 5.31 (s, 2H), 5.14 (m, 1H), 4.53 (m, 1H),4.35 (m, 1H), 4.03 (m, 1H), 3.49 (m, 4H), 3.24 (m, 1H), 2.98 (m, 3H),2.81 (m, 1H), 2.62 (m, 1H), 2.41 (s, 3H), 2.35 (m, 1H), 1.18 (t, J=7.6Hz, 3H)

Example 128:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-2-(4-methylpiperazin-1-yl)pyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Step 1:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-2-(methylthio)pyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

The glass was flame dried in vacuo and backfilled with argon. Then thedry argon flushed flask was charged with5-hydroxy-2-(methylthio)pyrimidine-4-carboxylic acid (194 mg, 767 μmol)and anhydrous THE (10.2 mL). The solution mixture was treated withpyridine (418 μL, 5.11 mmol) and cooled at 0° C. HOBT (237 mg, 1.53mmol) was added prior to EDC.HCl (300 mg, 1.53 mmol) and the resultingmixture was stirred for 15 minutes.2-(2-(3,6-Dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate AP) (300 mg, 511 μmol) was added. The bath was removed,and the resulting mixture was stirred at RT for 5 days. The reaction wasquenched with aq 0.35% NaHCO₃ (30 mL). DCM (30 mL) was added. Theorganic layer was collected, and the aqueous layer was back-extractedwith DCM (3×20 mL). The combined organic layers were washed with brine(60 mL), dried through a phase separator, and concentrated under reducedpressure to give a yellow oil (639 mg). The crude product was adsorbedonto Isolute and purified by column chromatography (Flash Pure ID HPsilica cartridge 24 g, eluent DCM:DCM/MeOH (8/2) 100:0 to 70:30). Theproduct containing fractions were combined and concentrated underreduced pressure to give the title compound as a white solid (203 mg,98% pure, yield: 55%).

LC-MS: Rt=1.07 min; MS m/z [M+H]⁺ 714.3, m/z [M−H]⁻ 712.3; UPLC-MS 1

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-2-(methylsulfinyl)pyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

A dry argon flushed flask was charged with2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-2-(methylthio)pyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(200 mg, 275 μmol) and anhydrous DCM (2.75 mL). The clear solution wascooled to 0-5° C. and m-chloroperbenzoic acid (61.5 mg, 275 μmol) wasadded. The RM was stirred under ice cooling for 1.75 hours. The RM wasquenched with aq 0.35% NaHCO₃ (25 mL). DCM (15 mL) was added. Theorganic layer was collected, and the aqueous layer was back-extractedwith DCM (3×15 mL). The combined organic layers were washed with brine(40 mL), dried through a phase separator, and concentrated under reducedpressure to give the title compound as a white powder (152 mg, 97% pure,yield: 74%).

LC-MS: Rt=0.89 min; MS m/z [M+H]⁺ 730.3, m/z [M−H]⁻ 728.3; UPLC-MS 1

Step 3:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-2-(4-methylpiperazin-1-yl)pyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

The vial was charged with2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-2-(methylsulfinyl)pyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(105 mg, 140 μmol), anhydrous NMP (1.4 mL) and 1-methylpiperazine (157μL, 1.40 mmol). The reaction tube was sealed and irradiated at 200° C.for 1.5 hours. The RM was filtered and purified by reverse phasepreparative HPLC (RP-HPLC acidic 1: 20 to 45% B in 15 min, RP-HPLCacidic 1: 20 to 40% B in 15 min with a plateau at 55% for 1 min). Theproduct containing fractions were combined and lyophilized to give thetitle compound as a beige solid (4.10 mg, 98% pure, yield: 3%).

LC-MS: Rt=0.87 min; MS m/z [M+H]⁺ 766.4, m/z [M−H]⁻ 764.5; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.02 (s, 1H), 9.80 (s, 1H), 8.20 (s, 1H),7.71 (d, J=8.8 Hz, 1H), 7.63 (m, 1H), 7.53 (d, J=8.8 Hz, 1H), 6.83 (m,1H), 5.25 (s, 2H), 4.49 (m, 3H), 4.26 (m, 2H), 3.81 (m, 2H), 3.44 (m,5H), 3.26 (m, 3H), 3.00 (m, 5H), 2.82 (s, 3H), 2.79 (m, 1H), 2.65 (m,1H), 2.53 (m, 2H), 2.35 (s, 3H), 1.20 (t, J=7.3 Hz, 3H)

Example 129:2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-5-methyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

2-(2-(3,6-Dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide(Intermediate ES) (172 mg, 232 μmol) was dissolved in DCM (4 mL) at 0°C. under argon. 3-Hydroxypicolinoyl chloride (Intermediate CV) (54.8 mg,348 μmol) was added slowly, followed by DIPEA (202 μL, 1.16 mmol). TheRM was stirred at 0° C. for 20 minutes, the at RT for 1.5 hours.3-Hydroxypicolinoyl chloride (Intermediate CV) (30.0 mg, 191 μmol) wasadded slowly, followed by DIPEA (100 μL, 574 μmol). The RM was stirredat RT for 30 minutes. Water (5 mL) and aq sat NaHCO₃ (5 mL) were addedand the mixture was extracted with DCM (4×50 mL). The combined organiclayers were washed with aq sat NaHCO₃ and water, dried through a phaseseparator and concentrated under reduced pressure. The crude product waspurified by reverse phase preparative HPLC (RP-HPLC acidic 1: 15 to 85%B in 20 min with a plateau at 95% for 1 min). The product containingfractions were combined, basified with aq sat NaHCO₃, ACN was removedand the residue was extracted with DCM (4×25 mL). The combined organiclayers were washed with water (10 mL), dried through a phase separatorand concentrated under reduced pressure to give the title compound as apale grey solid (99.1 mg, 100% pure, yield: 65%).

LC-MS: Rt=0.86 min; MS m/z [M+H]⁺ 654.4, m/z [M−H]⁻ 652.4; UPLC-MS 4

¹H NMR (600 MHz, DMSO-d₆) δ 10.39 (s, 1H), 10.25 (s, 1H), 8.23 (d, J=8.3Hz, 1H), 8.07 (m, 1H), 7.74 (d, J=8.4 Hz, 1H), 7.29 (m, 2H), 6.84 (m,1H), 5.31 (s, 2H), 4.53 (m, 1H), 4.27 (m, 2H), 3.81 (m, 2H), 3.45 (m,2H), 3.39 (m, 1H), 3.24 (m, 1H), 2.98 (m, 1H), 2.78 (m, 1H), 2.62 (m,1H), 2.60 (s, 3H), 2.58 (s, 3H), 2.54 (m, 2H)

Example 130: tert-butyl4-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate

To a stirred solution of2-(2-bromo-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate ET) (100 mg, 113 μmol), K₃PO₄ (72.3 mg, 340 μmol) andPdCl2(dppf).DCM adduct (9.27 mg, 11.0 μmol) in 1,4-dioxane (4 mL) andwater (2 mL) was added at RT tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate(52.6 mg, 170 μmol) and the RM was stirred at 80° C. for 15 minutes. TheRM was diluted with EtOAc and water, extracted twice with EtOAc and thecombined organic layers were washed with brine, dried over Na₂SO₄ andconcentrated under reduced pressure. The crude product was firstpurified by column chromatography (RediSep Column: Silica 12 g, eluentDCM:DCM/MeOH (8/2) 100:0 to 40:60), then by reverse phase preparativeHPLC (RP-HPLC basic 1: 15 to 95% B in 20 min). The product containingfractions were lyophilized to give the title compound as a white solid(62.0 mg, 98% pure, yield: 69%).

LC-MS: Rt=1.26 min; MS m/z [M+H]⁺ 781.6; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.02 (m, 2H), 8.56 (s, 1H), 7.73 (d, J=8.2Hz, 1H), 7.64 (m, 1H), 7.54 (d, J=8.5 Hz, 1H), 6.79 (m, 1H), 5.25 (s,2H), 4.52 (m, 1H), 4.06 (m, 2H), 3.53 (m, 3H), 3.47 (m, 2H), 3.23 (m,1H), 3.00 (m, 3H), 2.83 (m, 1H), 2.66 (m, 1H), 2.54 (m, 2H), 2.44 (s,3H), 2.36 (s, 3H), 1.43 (s, 9H), 1.20 (t, J=7.2 Hz, 3H)

Example 131:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((1S,6R)-5-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideorN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((1R,6S)-5-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-((1S,6R)-5-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamideor2-(6-((1R,6S)-5-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

2-(6-((1S,6R)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamideor2-(6-((1R,6S)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(Intermediate EX) (38.0 mg, 58.0 μmol),5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (Intermediate CY)(16.0 mg, 64.0 μmol) and HATU (26.0 mg, 69.0 μmol) were mixed in DMF (1mL). DIPEA (20.0 μL, 120 μmol) was added and the RM was stirred at RTfor 5 minutes. The RM was diluted with EtOAc and water, extracted twicewith EtOAc and the combined organic layers were washed with brine, driedover Na₂SO₄ and concentrated under reduced pressure. The crude productwas purified by column chromatography (RediSep Column: Silica 12 g,eluent DCM:DCM/MeOH (8/2) 100:0 to 50:50). The product containingfractions were combined and concentrated under reduced pressure to givethe title compound as a white solid (25.0 mg, 94% pure, yield: 50%).

LC-MS: Rt=1.28 min; MS m/z [M+H]⁺ 818.5/820.1, m/z [M−H]⁻ 816.4/818.4;UPLC-MS 1

LC-MS: Rt=6.07 min; MS m/z [M+H]⁺ 818.6/820.1, m/z [M−H]⁻ 816.3/818.4;UPLC-MS 2

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((1S,6R)-5-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideorN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((1R,6S)-5-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

2-(6-((1S,6R)-5-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamideor2-(6-((1R,6S)-5-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(25.0 mg, 31.0 μmol) was mixed with TFA (500 μL, 6.00 mmol) and the RMwas stirred at RT for 3 days. The RM was diluted with DCM and NaHCO₃,extracted twice with DCM and the combined organic layers were washedwith brine, dried over Na₂SO₄, and concentrated under reduced pressure.The crude product was purified by reverse phase preparative HPLC(RP-HPLC acidic 1: 5 to 95% B in 20 min) to give the title compound as awhite solid (14.0 mg, 100% pure, yield: 63%).

LC-MS: Rt=1.11 min; MS m/z [M+H]⁺ 728.4/730.4, m/z [M−H]⁻ 726.3/728.3;UPLC-MS 1

LC-MS: Rt=5.49 min; MS m/z [M+H]⁺ 728.4/730.4, m/z [M−H]⁻ 726.4/728.3;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ

Example 132:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((1S,6R)-5-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideorN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((1R,6S)-5-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-((1S,6R)-5-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamideor2-(6-((1R,6S)-5-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

2-(6-((1S,6R)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamideor2-(6-((1R,6S)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(Intermediate EY) (38.0 mg, 64.0 μmol),5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (Intermediate CY)(17.0 mg, 71.0 μmol) and HATU (29.0 mg, 77.0 μmol) were mixed in DMF (1mL). DIPEA (22.0 μL, 130 μmol) was added and the RM was stirred at RTfor 5 minutes. The RM was diluted with EtOAc and water, extracted twicewith EtOAc and the combined organic layers were washed with brine, driedover Na₂SO₄ and concentrated under reduced pressure. The crude productwas purified by column chromatography (RediSep Column: Silica 12 g,eluent DCM:DCM/MeOH (8/2) 100:0 to 50:50). The product containingfractions were combined and concentrated under reduced pressure to givethe title compound as a white solid (25.0 mg, 98% pure, yield: 47%).

LC-MS: Rt=1.27 min; MS m/z [M+H]⁺ 818.5/820.5, m/z [M−H]⁻ 816.5/818.6;UPLC-MS 1

LC-MS: Rt=6.14 min; MS m/z [M+H]⁺ 818.4/820.4, m/z [M−H]⁻ 816.4/818.4;UPLC-MS 2

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((1S,6R)-5-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideorN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((1R,6S)-5-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

2-(6-((1S,6R)-5-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamideor2-(6-((1R,6S)-5-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(25.0 mg, 31.0 μmol) was mixed with TFA (500 μL, 6.00 mmol) and the RMwas stirred at RT for 3 days. The RM was diluted with DCM and NaHCO₃,extracted twice with DCM and the combined organic layers were washedwith brine, dried over Na₂SO₄, and concentrated under reduced pressure.The crude product was purified by reverse phase preparative HPLC(RP-HPLC acidic 1: 5 to 95% B in 20 min) to give the title compound as awhite solid (16.0 mg, 100% pure, yield: 72%).

LC-MS: Rt=1.13 min; MS m/z [M+H]⁺ 728.5/730.5, m/z [M−H]⁻ 726.5/728.5;UPLC-MS 1

LC-MS: Rt=5.50 min; MS m/z [M+H]⁺ 728.4/730.4, m/z [M−H]⁻ 726.4/728.4;UPLC-MS 2

¹H NMR (600 MHz, DMSO-d₆) δ 10.4 (m, 2H), 8.56 (d, J=11.1 Hz, 1H), 8.04(d, J=9.3 Hz, 1H), 7.6 (m, 1H), 7.71 (d, J=8.3 Hz, 1H), 6.83 (m, 1H),5.45 (m, 1H), 5.23 (m, 1H), 4.95 (m, 1H), 4.36 (m, 1H), 4.24 (m, 2H),4.13 (m, 1H), 3.99 (m, 1H), 3.79 (m, 2H), 3.57 (m, 1H), 2.89 (m, 1H),2.71 (m, 3H), 2.50 (m, 2H), 2.44 (d, broad, J=13.6 Hz, 3H), 1.96 (m,1H), 1.41 (m, 1H), 1.21 (m, 4H)

Example 133:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl-2,2,3,3,5,5,6,6-d8)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To 5-hydroxy-6-methylpyrimidine-4-carboxylic acid (102 mg, 665 μmol) inDCM (3 mL) was added 1-chloro-N,N,2-trimethylprop-1-en-1-amine (114 μL,859 μmol). The RM was stirred at RT for 1 hour.N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl-2,2,3,3,5,5,6,6-d8)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate EZ) (318 mg, 554 μmol) and DCM (1 mL) were added. The RMwas stirred at RT for 30 minutes. DIPEA (290 μL, 1.66 mmol) was addeddropwise and the mixture was continued stirring at RT for 2.5 hours. TheRM was diluted with 0.5M HCl (1.6 mL) and DCM (5 mL). The biphasicmixture was stirred vigorously for 1 minute. The organic layer wasseparated. The aqueous layer was extracted with DCM (5 mL). The combinedorganic layers were dried through a phase separator and concentratedunder reduced pressure. The crude product was purified in 2 portions byreverse phase preparative HPLC (RP-HPLC acidic 1: 20 to 75% B in 20min), (RP-HPLC acidic 1: 25 to 54% B in 20 min). The product containingfractions were combined, basified to pH 8 with aq sat NaHCO₃, extractedwith DCM (2×100 mL), dried over Na₂SO₄ and concentrated under reducedpressure to give the title compound as a colourless solid (174 mg, 99%pure, yield: 44%).

LC-MS: Rt=1.03 min; MS m/z [M+H]⁺ 710.4/712.4, m/z [M−H]⁻ 708.4/710.4;UPLC-MS 1

LC-MS: Rt=5.12 min; MS m/z [M+H]⁺ 710.3/712.3, m/z [M−H]⁻ 708.4/710.3;UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 10.37 (m, 2H), 8.57 (s, 1H), 8.06 (d, J=8.5Hz, 1H), 7.97 (m, 1H), 7.72 (dd, J=2.2 Hz, 9.2 Hz, 1H), 6.84 (m, 1H),5.32 (s, 2H), 4.26 (m, 2H), 3.81 (m, 2H), 2.99 (m, 2H), 2.52 (m, 2H),2.45 (s, 3H), 1.19 (t, J=7.4 Hz, 3H)

Example 134:2-(5-cyclopropyl-2-(3,6-dihydro-2H-pyran-4-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

3-Hydroxypicolinic acid (49.1 mg, 353 μmol) was mixed with DCM (1.85 mL)at RT under argon. 1-Chloro-N,N,2-trimethylprop-1-en-1-amine (51.9 mg,389 μmol) was added and the RM was stirred at RT for 1.75 hours. Thissuspension was added to a cooled suspension of2-(5-cyclopropyl-2-(3,6-dihydro-2H-pyran-4-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide(Intermediate FA) (96.0 mg, 177 μmol) in DCM (1.5 mL) and DIPEA (154 μL,883 μmol). The resulting brown solution was stirred at RT for 2.5 hours.3-Hydroxypicolinic acid (32.4 mg, 233 μmol) was mixed with DCM (1.22 mL)at RT under argon. 1-Chloro-N,N,2-trimethylprop-1-en-1-amine (34.3 mg,257 μmol) was added and the RM was stirred at RT for 1.75 hours. Thissolution was added to the previous solution, followed by DIPEA (105 μL,602 μmol). The RM was stirred at RT for 1 hour. The RM was quenched withwater (4 mL) and aq sat NaHCO₃ (3 mL), then it was extracted with DCM(4×20 mL). The organic layer was washed with aq sat NaHCO₃ and water.The organic layer was dried through a phase separator and concentratedunder reduced pressure. The aqueous layer was extracted with EtOAc (3×40mL). The organic layer was dried through a phase separator, combinedwith the first crude and concentrated under reduced pressure. The crudeproduct (140 mg) was purified by reverse phase preparative HPLC (RP-HPLCacidic 1: 25 to 75% in 20 min). The product containing fractions werecombined, basified with aq sat NaHCO₃ and ACN was removed under reducedpressure. The aqueous residue was extracted with DCM (4×30 mL) andwater. The organic layer was dried through a phase separator andconcentrated under reduced pressure to give the title compound as awhite solid (13.7 mg). The impure product containing fractions werecombined and purified again by reverse phase preparative HPLC (RP-HPLCacidic 1: 25 to 75% in 20 min). The product containing fractions wereneutralized trough a PL-HCO3 MP SPE (500 mg per 6 mL) cartridge to givethe title compound as a white solid (5.90 mg). Both batches werecombined to give the title compound as a white solid (19.6 mg, 99% pure,yield: 17%).

LC-MS: Rt=0.99 min; MS m/z [M+H]⁺ 665.4, m/z [M−H]⁻ 663.4; UPLC-MS 3

¹H NMR (600 MHz, DMSO-d₆) δ 10.86 (s, 1H), 10.39 (s, 1H), 8.07 (m, 1H),7.78 (m, 2H), 7.71 (m, 2H), 7.30 (m, 2H), 6.80 (m, 1H), 5.35 (s, 2H),4.40 (m, 1H), 4.24 (m, 2H), 3.80 (m, 2H), 3.42 (m, 3H), 3.25 (m, 1H),3.10 (m, 1H), 2.87 (m, 1H), 2.71 (m, 1H), 2.58 (m, 2H), 1.76 (m, 1H),1.24 (m, 4H)

Example 135:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-(2-hydroxyethyl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-(2-hydroxyethyl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate FB) (152 mg, 76.0 μmol) and 3-hydroxypicolinoyl chloride(Intermediate CV) (13.1 mg, 83.0 μmol) were dissolved in DCM (1 mL) andDIPEA (26.0 μL, 151 μmol) was added. The mixture was stirred at RT for 1day. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL) were added.The aqueous layer was washed with DCM (2×10 mL). The combined organiclayers were dried through a phase separator and concentrated underreduced pressure. The crude product was purified by columnchromatography (RediSep Column: Silica 12 g, eluent DCM:DCM/MeOH (8/2)100:0 to 60:40). The product containing fractions were combined,concentrated under vacuum, and purified by reverse phase preparativeHPLC (RP-HPLC acidic 1: 5 to 100% B in 20 min). The product containingfractions were combined, basified with aq sat NaHCO₃, extracted twicewith DCM, dried through a phase separator, and concentrated underreduced pressure to give the title compound (17.2 mg, 97% pure, yield:32%).

LC-MS: Rt=0.92 min; MS m/z [M+H]⁺ 683.5, m/z [M−H]⁻ 681.5; UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.37 (s, 1H), 10.01 (s, 1H), 8.06 (m, 1H),7.73 (d, J=8.5 Hz, 1H), 7.62 (m, 1H), 7.53 (d, J=8.5 Hz, 1H), 7.28 (m,2H), 6.84 (m, 1H), 5.34 (s, 2H), 5.02 (m, 1H), 4.53 (m, 1H), 4.26 (m,2H), 3.81 (m, 2H), 3.73 (m, 2H), 3.44 (m, 3H), 3.20 (m, 3H), 2.95 (m,1H), 2.81 (m, 1H), 2.64 (m, 1H), 2.51 (m, 2H), 2.35 (s, 3H)

Example 136:2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-(1-(3-hydroxypicolinoyl)piperidin-4-yl)-5-methyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

3-Hydroxypicolinic acid (8.89 mg, 64.0 μmol) was mixed with thionylchloride (6.91 mg, 58.0 μmol) at 80° C. for 1 hour. S02 gas evolutionwas detected. The RM was cooled down, dissolved with toluene, andevaporated to dryness. The residue was dissolved in DCM (1.16 mL) andDIPEA (40.6 μL, 232 μmol) was added, followed by2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-6-(piperidin-4-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide(Intermediate FC) (30.0 mg, 58.0 μmol). The RM was stirred at RT for 2days. DCM was added and the RM was washed with aq sat NaHCO₃ and water.The organic layer was dried through a phase separator and concentratedunder reduced pressure. The crude product was purified by columnchromatography (Silica gel column, eluent DCM:MeOH 100:0 to 95:5), thenby reverse phase preparative HPLC (RP-HPLC basic 1: 6 to 36% B in 20min) to give the title compound (2.00 mg, 90% pure, yield: 5%).

LC-MS: Rt=0.95 min; MS m/z [M+H]⁺ 638.2, m/z [M−H]⁻ 636.2; UPLC-MS 4

LC-MS: Rt=4.55 min; MS m/z [M+H]⁺ 638.2, m/z [M−H]⁻ 636.2; UPLC-MS 14

¹H NMR (600 MHz, DMSO-d₆) δ 10.91 (m, 2H), 8.00 (m, 1H), 7.78 (d, J=8.4Hz, 2H), 7.70 (d, J=8.6 Hz, 2H), 7.24 (m, 2H), 6.80 (m, 1H), 5.25 (s,2H), 4.64 (m, 1H), 4.23 (m, 2H), 3.79 (m, 2H), 3.47 (m, 1H), 3.12 (m,3H), 2.82 (m, 1H), 2.50 (m, 5H), 2.31 (m, 1H), 1.59 (m, 1H), 1.41 (m,1H)

Example 137N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-((3-hydroxypyridin-2-yl)sulfonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Step 1:2-(6-(4-((3-(benzyloxy)pyridin-2-yl)sulfonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

To a stirred solution ofN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(Intermediate AK) (71 mg, 125 μmol) was added triethylamine (35 μL, 250mmol) followed by (3-(benzyloxy)pyridine-2-sulfonyl chloride(commercially available) (36 mg, 125 μmol) at RT and the RM was stirredat RT for 10 minutes. Diluted with water (1 ml) and DCM (5 ml) andstirred vigorously for 1 minute. The organic phase was separated byfiltering through a phase separator and concentrated under reducedpressure to afford the title compound as a pale brown solid.

LC-MS: Rt=1.25 min; MS m/z [M+H]⁺ 813.5/815.5, m/z [M−H]⁻ 811.5/813.5;UPLC-MS 1

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-((3-hydroxypyridin-2-yl)sulfonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

2-(6-(4-((3-(benzyloxy)pyridin-2-yl)sulfonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamidewas mixed with TFA (2 ml) and stirred at 60° C. for 18 hours. The RM wasconcentrated under reduced pressure and purified by reverse phasepreparative HPLC (RP-HPLC acidic 1: 20 to 60% B in 20 min). The productcontaining fractions were combined, basified with aq sat NaHCO₃ and theACN was removed under reduced pressure. The residue was extracted withDCM (3×10 mL), dried over Na₂SO₄ and concentrated under reduced pressureto give the title compound as a colourless solid.

LC-MS: Rt=1.08 min; MS m/z [M+H]⁺ 723.1/725.0, m/z [M−H]⁻ 721.1/723.1;UPLC-MS 1

¹H NMR (400 MHz, DMSO-d₆) δ 10.97 (s, 1H), 10.37 (s, 1H), 8.14 (m, 1H),8.06 (m, 1H), 7.98 (m, 1H), 7.72 (m, 1H), 7.51 (m, 2H), 6.83 (m, 1H),5.32 (s, 2H), 4.27 (m, 2H), 3.79 (m, 3H), 3.56 (m, 2H), 3.19 (m, 3H),2.94 (m, 2H), 2.74 (m, 2H), 1.17 (m, 3H)

Preparation of Intermediates

Intermediate A: tert-butyl4-(5-ethyl-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To the stirred solution of 3-morpholino-1H-1,2,4-triazol-5-amine (12.0g, 71.0 mmol) in EtOH (100 mL), were added tert-butyl4-(1-ethoxy-1,3-dioxopentan-2-yl)piperazine-1-carboxylate (IntermediateEJ) (34.9 g, 106 mmol) and H₃PO₄ (3.70 mL, 71.0 mmol) at RT. The RM wasstirred at 110° C. for 48 hours. The RM was filtered to obtain a firstportion of the product. To the filtrate were added Boc₂O (16.5 mL, 71.0mmol) and DIPEA (12.4 mL, 71.0 mmol) at 0° C. and the RM was stirred atRT for 16 hours. The RM was filtered to obtain a second portion of theproduct which was combined with the first portion to give the titlecompound.

LC-MS: Rt=1.43 min; MS m/z [M+H]⁺ 434.2; UPLC-MS 11

Intermediate B:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To tert-butyl4-(5-ethyl-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate A) (500 mg, 1.15 mmol) in DMF (4 mL) was added DIPEA (504μL, 2.88 mmol) followed byN-(2-chloro-4-(trifluoromethyl)phenyl)-2-iodoacetamide (Intermediate DL)(440 mg, 1.21 mmol). The RM was stirred at RT for 24 hours. The RM wasdiluted with water (10 mL) and the resulting suspension was stirred atRT for 20 minutes, filtered and washed with water (10 mL) then driedunder vacuum to give the title compound as an off-white solid.

LC-MS: Rt=1.30 min; MS m/z [M−H]⁻ 667.5/669.5; UPLC-MS 1

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(700 mg, 1.05 mmol) was dissolved in DCM (5 mL) and TFA (1.21 mL, 15.7mmol) was added and the RM was stood at RT for 2 hours. The RM wasevaporated in vacuo to give the desired product trifluoroacetate salt asa brown solid.

LC-MS: Rt=0.78 min; MS m/z [M+H]⁺ 569.3/571.3, m/z [M−H]⁻ 567.2/569.2;UPLC-MS 1

The free base of intermediate B can be prepared by partitioning the TFAsalt between dichloromethane and saturated aqueous NaHCO₃, followed byseparation and drying of the organic layer.

Intermediate C:N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((5-chloro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To the stirred solution of tert-butyl4-(5-ethyl-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate A) (600 mg, 1.38 mmol) in DMF (4 mL) were added2-bromo-N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate DM) (550 mg, 1.66 mmol) and DIPEA (600 μL, 3.47 mmol) at0° C. The RM was stirred at RT for 16 hours. The RM was concentratedunder reduced pressure and ice was added to the remaining suspension,then it was filtered and dried under reduced pressure. The crude productwas purified by column chromatography (Silica gel column: Silica 12 g,eluent DCM:MeOH 95:5 to 90:10). The product containing fractions werecombined and concentrated under reduced pressure to give the titlecompound as a brown solid.

LC-MS: Rt=1.64 min; MS m/z [M−H]⁻ 681.3/683.3; UPLC-MS 11

Step 2:N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To tert-butyl4-(4-(2-((5-chloro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(1.00 g, 1.46 mmol) was added TFA (10.0 mL, 130 mmol) and DCM (10 mL) at0° C. The RM was stirred at 40° C. for 14 hours. The RM was concentratedunder reduced pressure to give the title compound.

LC-MS: Rt=1.36 min; MS m/z [M+H]⁺ 583.2/585.2; UPLC-MS 11

Intermediate D:2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(5-ethyl-4-(2-((5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(5-ethyl-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate A) (350 mg, 807 μmol) was suspended in DMF (4 mL).2-Bromo-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate DN) (254 mg, 807 μmol) and DIPEA (423 μL, 2.42 mmol) wereadded at 0° C. and the RM was stirred at RT for 12 hours. The RM wascooled to 0° C., water was added and the precipitate was filtered,washed with hexane and dried under reduced pressure. The crude productwas purified by column chromatography (Silica gel column: Silica 12 g,eluent DCM:MeOH 100:0 to 98:2). The product containing fractions werecombined, concentrated and dried under HV to give the title compound asan off-white solid.

LC-MS: Rt=1.63 min; MS m/z [M+H]⁺ 667.3; UPLC-MS 11

Step 2:2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

To the solution of tert-butyl4-(5-ethyl-4-(2-((5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(300 mg, 449 μmol) in 1,4-dioxane (10 mL) was added HCl 4M in1,4-dioxane (10.0 mL, 40.0 mmol) at 0° C. and the RM was stirred at RTfor 16 hours. The RM was concentrated under reduced pressure and washedwith Et₂O (2×10 mL) and dried under reduced pressure to give the HClsalt of the title compound as an off-white solid.

LC-MS: Rt=1.35 min; MS m/z [M+H]⁺ 567.3; UPLC-MS 11

Intermediate E:N-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(5-ethyl-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate A) (400 mg, 920 μmol) was suspended in DMF (5 mL).2-Bromo-N-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)acetamide(Intermediate DO) (463 mg, 1.38 mmol) and DIPEA (403 μL, 2.31 mmol) wereadded at 0° C. and the RM was stirred at RT for 14 hours. The RM wascooled to 0° C., water was added and the precipitate was filtered off,washed with hexane and dried under reduced pressure. The crude productwas purified by column chromatography (Silica gel column: Silica 12 g,eluent DCM:MeOH 100:0 to 98:2). The product containing fractions werecombined, concentrated under reduced pressure and dried under HV to givethe title compound.

LC-MS: Rt=1.67 min; MS m/z [M+H]⁺ 687.3/689.3; UPLC-MS 11

Step 2:N-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To the solution of tert-butyl4-(4-(2-((2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(400 mg, 582 μmol) in DCM (5 mL) was added HCl 4M in 1,4-dioxane (10.0mL, 40.0 mmol) at 0° C. and the reaction mixture was stirred at RT for16 hours. The RM was concentrated under reduced pressure, washed withEt₂O (2×10 mL) and dried under HV to give the title compound as anoff-white solid, as the HCl salt.

LC-MS: Rt=1.34 min; MS m/z [M+H]⁺ 587.2/589.2; UPLC-MS 11

Intermediate F:2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To the stirred solution of tert-butyl4-(5-ethyl-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate A) (500 mg, 1.15 mmol) in DMF (5 mL) were added2-bromo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (IntermediateDP) (410 mg, 1.38 mmol) and DIPEA (504 μL, 2.88 mmol). The RM wasstirred at RT for 16 hours. The RM was concentrated under reducedpressure, water was added and the RM was extracted with DCM. Thecombined organic layers were dried over Na₂SO₄, concentrated and driedunder reduced pressure. The crude product was purified by columnchromatography (Silica gel column: Silica 12 g, eluent DCM:MeOH 95:5 to90:10). The product containing fractions were combined and concentratedunder reduced pressure to give the title compound as a brown stickyliquid.

LC-MS: Rt=1.65 min; MS m/z [M+H]⁺ 649.3; UPLC-MS 11

Step 2:2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To the stirred solution of tert-butyl4-(5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(500 mg, 771 μmol) in 1,4-dioxane (15 mL) was added HCl 4M in1,4-dioxane (7.50 mL, 30.0 mmol) at 0° C. The RM was stirred at RT for12 hours. The RM was concentrated, washed three times with a mixture ofpentane and Et₂O (1:1) and dried under reduced pressure to give thetitle compound, as the HCl salt.

LC-MS: Rt=1.35 min; MS m/z [M+H]⁺ 549.2; UPLC-MS 11

Intermediate G:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((2-chloro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-5-ethyl-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(5-ethyl-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate A) (950 mg, 2.19 mmol) was suspended in DMF (5 mL).2-Bromo-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide(Intermediate DQ) (626 mg, 1.97 mmol) and DIPEA (957 μL, 5.48 mmol) wereadded at 0° C. and the RM was stirred at RT for 12 hours. Water wasadded and the precipitate was filtered off, washed with Et₂O and driedunder HV to give the title compound.

LC-MS: Rt=1.81 min; MS m/z [M+H]⁺ 670.0/672.0; UPLC-MS 12

Step 2:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((2-chloro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-5-ethyl-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(1.10 g, 1.36 mmol) was suspended in 1,4-dioxane (10 mL) and HCl 4M in1,4-dioxane (10.0 mL, 40.0 mmol) was added at 0° C. and the RM wasstirred at RT for 12 hours. The RM was concentrated under reducedpressure to give the title compound, as the HCl salt.

LC-MS: Rt=1.35 min; MS m/z [M+H]⁺ 570.2/572.2; UPLC-MS 11

Intermediate H:2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(5-ethyl-4-(2-((2-fluoro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To the stirred solution of tert-butyl4-(5-ethyl-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate A) (800 mg, 1.85 mmol) in DMF (6 mL) were added2-bromo-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide (IntermediateDR) (665 mg, 2.22 mmol) and DIPEA (806 μL, 4.61 mmol) at 0° C. and theRM was stirred at RT for 16 hours. The RM was concentrated under reducedpressure, ice cold water was added and the precipitate was filtered offand dried under reduced pressure. The crude product was purified bycolumn chromatography (Silica gel column: Silica 24 g, eluent DCM:MeOH100:0 to 90:10). The product containing fractions were combined,concentrated under reduced pressure and dried under HV to give the titlecompound as a brown solid.

LC-MS: Rt=1.60 min; MS m/z [M−H]⁻ 651.3; UPLC-MS 11

Step 2:2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide

To the stirred solution of tert-butyl4-(5-ethyl-4-(2-((2-fluoro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(500 mg, 766 μmol) in DCM (10 mL) was added HCl 4M in 1,4-dioxane (2.00mL, 8.00 mmol) at 0° C. and the RM was stirred at RT for 6 hours. The RMwas concentrated under reduced pressure to give the title compound, asthe HCl salt.

LC-MS: Rt=1.32 min; MS m/z [M+H]⁺ 553.3; UPLC-MS 11

Intermediate I:N-(4-chloro-2-methyl-5-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((4-chloro-2-methyl-5-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(5-ethyl-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate A) (600 mg, 1.38 mmol) was suspended in DMF (10 mL).2-Bromo-N-(4-chloro-2-methyl-5-(trifluoromethyl)phenyl)acetamide(Intermediate DS) (503 mg, 1.52 mmol) and DIPEA (604 μL, 3.46 mmol) wereadded at 0° C. and the RM was stirred at RT for 12 hours. The RM wascooled to 0° C., water was added, the precipitate was filtered off,washed with hexane and dried under reduced pressure. The crude productwas purified twice by column chromatography (Silica gel column: Silica12 g, eluent DCM:MeOH 100:0 to 98:2). The product containing fractionswere combined, concentrated and dried under HV to give the titlecompound as an off-white solid.

LC-MS: Rt=1.64 min; MS m/z [M+H]⁺ 683.2/685.2; UPLC-MS 11

Step 2:N-(4-chloro-2-methyl-5-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To the solution of tert-butyl4-(4-(2-((4-chloro-2-methyl-5-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-morpholino-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(700 mg, 799 μmol) in 1,4-dioxane (10 mL) was added HCl 4M in1,4-dioxane (10.0 mL, 40.0 mmol) at 0° C. and the RM was stirred at RTfor 16 hours. The RM was concentrated under reduced pressure, washedwith Et₂O (2×10 mL) and dried under reduced pressure to give the titlecompound as a HCl salt.

LC-MS: Rt=1.08 min; MS m/z [M+H]⁺ 583.1/585.1; UPLC-MS 13

Intermediate J: rac-tert-butyl4-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylateStep 1: rac-3-(3-fluorocyclohexyl)-1H-1,2,4-triazol-5-amine

The rac-3-fluoropiperidine.HCl (4.90 g, 35.1 mmol) was suspended in ACN(250 mL), then dimethyl cyanocarbonimidodithioate (5.20 g, 35.6 mmol)and DIPEA (6.25 mL, 35.8 mmol) were added and the RM was heated at 80°C. for 14 hours. Hydrazine hydrate (1.74 mL, 35.8 mmol) was added to theRM which was stirred at 80° C. for 14 hours. The RM was concentrated,water was added and it was extracted with 10% MeOH in DCM. The combinedorganic layers were dried over Na₂SO₄, filtered and concentrated underreduced pressure to give the title compound.

LC-MS: Rt=0.24 min; MS m/z [M+H]⁺ 185.1; UPLC-MS 11

Step 2: rac-tert-butyl4-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To the stirred solution ofrac-3-(3-fluorocyclohexyl)-1H-1,2,4-triazol-5-amine (2.10 g, 11.3 mmol)in EtOH (30.0 mL) were added tert-butyl4-(1-ethoxy-1,3-dioxopentan-2-yl)piperazine-1-carboxylate (IntermediateEJ) (5.59 g, 17.0 mmol) and H₃PO₄ (1.33 g, 13.6 mmol) at RT. The RM wasstirred at 90° C. for 18 hours. Boc₂O (3.95 mL, 17.0 mmol) and DIPEA(4.95 mL, 28.3 mmol) were added at 0° C. and the RM was stirred at RTfor 20 hours. The RM was cooled to RT, the suspension was filtered andwashed with EtOH. The filtrate was concentrated under reduced pressureand the residue was purified by column chromatography (Silica gelcolumn: Silica 24 g, eluent DCM:MeOH 100:0 to 98:2) to get the titlecompound.

LC-MS: Rt=1.50 min; MS m/z [M+H]⁺ 450.3; UPLC-MS 11

Intermediate K:rac-N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: rac-tert-butyl4-(4-(2-((5-chloro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Rac-tert-butyl4-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate J) (150 mg, 334 μmol) was suspended in DMF (2 mL).2-Bromo-N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate DM) (88.0 mg, 267 μmol) and DIPEA (117 μL, 667 μmol) wereadded at 0° C. and the RM was stirred at RT for 12 hours. The RM wascooled to 0° C., water was added and the precipitate was filtered off,washed with hexane and dried under reduced pressure. The crude productwas combined with another batch and purified twice by columnchromatography (2× Silica gel column: Silica 12 g, eluent DCM:MeOH 100:0to 98:2). The product containing fractions were combined, concentratedand dried under HV to give the title compound as an off-white solid.

LC-MS: Rt=1.68 min; MS m/z [M−H]⁻ 697.2/699.2; UPLC-MS 11

Step 2:rac-N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To the solution of rac-tert-butyl4-(4-(2-((5-chloro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(410 mg, 516 μmol) in 1,4-dioxane (10 mL) was added HCl 4M in1,4-dioxane (10.0 mL, 4.00 mmol) at 0° C. and the RM was stirred at RTfor 16 hours. The RM was concentrated under reduced pressure, washedwith Et₂O (2×10 mL) and dried under HV to give the title compound as aHCl salt.

LC-MS: Rt=1.37 min; MS m/z [M+H]⁺ 599.3/601.3; UPLC-MS 11

Intermediate L:rac-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: rac-tert-butyl4-(5-ethyl-2-(3-fluoropiperidin-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Rac-tert-butyl4-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate J) (500 mg, 1.11 mmol) was suspended in DMF (10 mL).2-bromo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (IntermediateDP) (329 mg, 1.11 mmol) and DIPEA (486 μL, 2.78 mmol) were added at 0°C. and the RM was stirred at RT for 12 hours. The RM was cooled to 0°C., water was added and the precipitate was filtered off, washed withhexane and dried under reduced pressure. The crude product was purifiedby column chromatography (Silica gel column: Silica 24 g, eluentDCM:MeOH 100:0 to 95:5). The product containing fractions were combined,concentrated down and dried under HV to give the title compound.

LC-MS: Rt=1.64 min; MS m/z [M+H]⁺ 665.3; UPLC-MS 11

Step 2:rac-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

The solution of rac-tert-butyl4-(5-ethyl-2-(3-fluoropiperidin-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(350 mg, 527 μmol) in 1,4-dioxane (10 mL) was added HCl 4M in1,4-dioxane (1.32 mL, 5.27 mmol) and the RM was stirred at RT for 16hours. The RM was concentrated under reduced pressure. The resultingsolid was triturated with Et₂O to give the title compound, as the HClsalt.

LC-MS: Rt=1.37 min; MS m/z [M+H]⁺ 565.3; UPLC-MS 11

Intermediate M:rac-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: rac-tert-butyl4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Rac-tert-butyl4-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate J) (400 mg, 890 μmol) was suspended in DMF (2 mL).2-Bromo-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (IntermediateDT) (225 mg, 710 μmol) and DIPEA (311 μL, 1.78 mmol) were added at 0° C.and the RM was stirred at RT for 12 hours. Water was added, theprecipitate was filtered off and dried under reduced pressure. The crudeproduct was purified by column chromatography (Silica gel column: Silica12 g, eluent DCM:MeOH 100:0 to 98:2). The product containing fractionswere combined, concentrated and dried under reduced pressure to give thetitle compound.

LC-MS: Rt=1.67 min; MS m/z [M−H]⁻ 683.3/685.3; UPLC-MS 11

Step 2:rac-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Rac-tert-butyl4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(200 mg, 292 μmol) was suspended in DCM (5 mL) and HCl 4M in 1,4-dioxane(3.00 mL, 99.0 mmol) was added at 0° C. and the RM was stirred at RT for2 hours. The RM was concentrated under reduced pressure to give thetitle compound, as the HCl salt.

LC-MS: Rt=1.38 min; MS m/z [M+H]⁺ 585.2/587.2; UPLC-MS 11

Intermediate N:rac-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: rac-tert-butyl4-(4-(2-((2-chloro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Rac-tert-butyl4-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate J) (500 mg, 1.11 mmol) was suspended in DMF (10 mL).2-Bromo-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide(Intermediate DQ) (353 mg, 1.11 mmol) and DIPEA (389 μL, 2.23 mmol) wereadded at 0° C. and the RM was stirred at RT for 12 hours. The RM wascooled to 0° C., water was added and the precipitate was filtered off,washed with hexane and dried under reduced pressure. The crude productwas purified by column chromatography (Silica gel column: Silica 12 g,eluent DCM:MeOH 100:0 to 98:2). The product containing fractions werecombined, concentrated under reduced pressure and dried under HV to givethe title compound.

LC-MS: Rt=1.62 min; MS m/z [M−H]⁻ 684.2/686.2; UPLC-MS 11

Step 2:rac-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To the solution of rac-tert-butyl4-(4-(2-((2-chloro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(100 mg, 109 μmol) was dissolved in 1,4-dioxane (5 mL) and HCl 4M in1,4-dioxane (5.00 mL, 20.0 mmol) was added at 0° C. and the RM wasstirred at RT for 16 hours. The RM was concentrated under reducedpressure, washed with Et₂O (2×10 mL) and dried under HV to give thetitle compound as a HCl salt as an off-white solid.

LC-MS: Rt=0.53 min; MS m/z [M+H]+ 586.4/588.4; UPLC-MS 12

Intermediate O: tert-butyl4-(5-ethyl-7-oxo-2-(pyrrolidin-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To the stirred solution of 5-(pyrrolidin-1-yl)-4H-1,2,4-triazol-3-amine(1.00 g, 6.53 mmol) in EtOH (30.0 mL) were added tert-butyl4-(1-methoxy-1,3-dioxopentan-2-yl)piperazine-1-carboxylate (IntermediateEH) (3.08 g, 9.79 mmol) and H₃PO₄ (340 μL, 6.53 mmol) at RT. The RM wasstirred at 90° C. for 64 hours. Boc₂O (1.51 mL, 6.53 mmol) and DIPEA(2.85 mL, 16.3 mmol) were added at 0° C. and RM was stirred at RT for 16hours. The RM was filtered and the obtained solid was dried underreduced pressure to give the title compound as a white solid.

LC-MS: Rt=1.63 min; MS m/z [M+H]⁺ 418.2; UPLC-MS 13

Intermediate P:2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(pyrrolidin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-2-(pyrrolidin-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(5-ethyl-7-oxo-2-(pyrrolidin-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate O) was suspended in DMF (10 mL).2-bromo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (IntermediateDP) (468 mg, 1.58 mmol) and DIPEA (627 μL, 3.59 mmol) were added at 0°C. and the RM was stirred at RT for 12 hours. The RM was cooled to 0°C., water was added and the precipitate was filtered off, washed withhexane and dried under reduced pressure. The crude product was purifiedtwice by column chromatography (Silica gel column: Silica 12 g, eluentDCM:MeOH 100:0 to 98:2). The product containing fractions were combined,concentrated and dried under HV to give the title compound as anoff-white solid.

LC-MS: Rt=1.64 min; MS m/z [M+H]+ 633.3; UPLC-MS 11

Step 2:2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(pyrrolidin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To the solution of tert-butyl4-(5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-2-(pyrrolidin-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(750 mg, 759 μmol) in 1,4-dioxane (10 mL) was added HCl 4M in1,4-dioxane (10.0 mL, 40.0 mmol) at 0° C. and the RM was stirred at RTfor 16 hours. The RM was concentrated under reduced pressure, washedwith Et₂O (2×10 mL) and dried under HV to give the title compound, asthe HCl salt.

LC-MS: Rt=1.34 min; MS m/z [M+H]+ 533.3; UPLC-MS 11

Intermediate Q:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(pyrrolidin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((2-chloro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-5-ethyl-7-oxo-2-(pyrrolidin-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To the stirred solution of tert-butyl4-(5-ethyl-7-oxo-2-(pyrrolidin-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate O) (700 mg, 1.68 mmol) in DMF (10 mL) were added2-bromo-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide(Intermediate DQ) (479 mg, 1.51 mmol) and DIPEA (732 μL, 4.19 mmol) at0° C. and the RM was stirred at RT for 16 hours. The RM was concentratedunder reduced pressure ice cold water was added and the precipitate wasfiltered off and dried under reduced pressure. The crude product waspurified by column chromatography (Silica gel column: Silica 24 g,eluent DCM:MeOH 100:0 to 90:10). The product containing fractions werecombined, concentrated and dried under HV to give the title compound asa brown solid.

LC-MS: Rt=1.64 min; MS m/z [M+H]+ 654.2/656.2; UPLC-MS 11

Step 2:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(pyrrolidin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To the stirred solution of tert-butyl4-(4-(2-((2-chloro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-5-ethyl-7-oxo-2-(pyrrolidin-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(500 mg, 680 μmol) in DCM (10 mL) was added HCl 4M in 1,4-dioxane (5.00mL, 20.0 mmol) at 0° C. and the RM was stirred at RT for 16 hours. TheRM was concentrated under reduced pressure to give the title compound,as the HCl salt.

LC-MS: Rt=1.37 min; MS m/z [M+H]+ 554.2/556.2; UPLC-MS 11

Intermediate R:N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(pyrrolidin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((5-chloro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-2-(pyrrolidin-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To the stirred solution of tert-butyl4-(5-ethyl-7-oxo-2-(pyrrolidin-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate O) (450 mg, 1.08 mmol) in DMF (5 mL) were added2-bromo-N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate DM) (356 mg, 1.08 mmol) and DIPEA (471 μL, 2.69 mmol) at0° C. and the RM was stirred at RT for 16 hours. The RM was concentratedunder reduced pressure, ice cold water was added, the precipitate wasfiltered off and dried under reduced pressure. The crude product waspurified by column chromatography (Silica gel column: Silica 24 g,eluent DCM:MeOH 100:0 to 90:10). The product containing fractions werecombined, concentrated and dried to give the title compound as a brownsolid.

LC-MS: Rt=1.67 min; MS m/z [M+H]+ 667.3/669.3; UPLC-MS 11

Step 2:N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(pyrrolidin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To the stirred solution of tert-butyl4-(4-(2-((5-chloro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-2-(pyrrolidin-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(200 mg, 300 μmol) in DCM (5 mL) was added HCl 4M in 1,4-dioxane (2.00mL, 8.00 mmol) at 0° C. and the RM was stirred at RT for 2 hours. The RMwas concentrated under reduced pressure to give the title compound, asthe HCl salt.

LC-MS: Rt=1.38 min; MS m/z [M+H]+ 567.2/569.2; UPLC-MS 11

Intermediate S:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(4-((2,2-difluoroethyl)(methyl)amino)piperidin-1-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep1:1-(5-amino-1H-1,2,4-triazol-3-yl)-N-(2,2-difluoroethyl)-N-methylpiperidin-4-amine

To N-(2,2-difluoroethyl)-N-methylpiperidin-4-amine (998 mg, 3.97 mmol)in ACN (10 mL) was added DIPEA (1.46 mL, 8.34 mmol) and the RM wasstirred at RT for 5 minutes. Then dimethyl cyanocarbonimidodithioate(581 mg, 3.97 mmol) was added and the RM was heated at 85° C. for 21hours. Hydrazine hydrate (328 μL, 6.76 mmol) was added and the RM wascontinued heating at 85° C. for 21 hours. Further hydrazine hydrate(85.0 μL, 1.75 mmol) was added and the RM was continued heating at 100°C. for 4 hours. Then it was allowed to cool to RT with stirring. Theresulting suspension was filtered and the solid was washed with ACN (10mL) and dried under vacuum to give the title compound as a colourlesssolid.

LC-MS: Rt=0.12 min; MS m/z [M+H]⁺ 261.3; UPLC-MS 1

Step 2: tert-butyl4-(2-(4-((2,2-difluoroethyl)(methyl)amino)piperidin-1-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To1-(5-amino-1H-1,2,4-triazol-3-yl)-N-(2,2-difluoroethyl)-N-methylpiperidin-4-amine(510 mg, 1.96 mmol) in EtOH (10 mL) was added H₃PO₄ (384 mg, 3.92 mmol)and the RM was stirred at RT for 3 minutes. Tert-butyl4-(1-methoxy-1,3-dioxopentan-2-yl)piperazine-1-carboxylate (IntermediateEH) (678 mg, 2.16 mmol) was added and the RM was heated at 85° C. for 18hours. While still warm, the orange solution was decanted from theresidual solid and evaporated in vacuo to give an orange gum which waspartitioned between DCM/MeOH (9/1, 30 mL) and 5% aq NaHCO₃ (20 mL). Theorganic layer was separated and the aqueous layer was extracted with DCM(2×20 mL). The combined organic layers were dried through a phaseseparator and concentrated under reduced pressure to give an orange gum.The crude product was purified in 2 portions by column chromatography(2×RediSep Column: Silica 12 g, eluent DCM:MeOH 100:0 to 92:08). Theproduct containing fractions were combined, concentrated and dried underHV to give the title compound as a dark pink solid.

LC-MS: Rt=0.76 min; MS m/z [M+H]⁺ 525.4/526.4, m/z [M−H]⁻ 523.5/524.5;UPLC-MS 1

Step 3: tert-butyl4-(4-(2-((2-chloro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-2-(4-((2,2-difluoroethyl)(methyl)amino)piperidin-1-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To tert-butyl4-(2-(4-((2,2-difluoroethyl)(methyl)amino)piperidin-1-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(162 mg, 309 μmol) in DMF (2 mL) was added DIPEA (81.0 μL, 463 μmol),followed by N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-iodoacetamide(Intermediate DU) (124 mg, 340 μmol). The RM was heated at 50° C. for 2hours.

LC-MS: Rt=1.11 min; MS m/z [M−H]⁻ 759.3/761.3; UPLC-MS 1

Step 4:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(4-((2,2-difluoroethyl)(methyl)amino)piperidin-1-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

The RM from step 3 was allowed to cool to RT then TFA (357 μL, 4.63mmol) was added and the RM was heated at 55° C. for 1 hour, then at 100°C. for 70 minutes. The RM was concentrated under reduced pressure togive a solution of desired product in DMF (1 mL). The solution waspartitioned between DCM (20 mL) and 5% aq NaHCO₃ (15 mL). The organiclayer was separated, dried through a phase separator and concentratedunder reduced pressure to give the title compound as a brown gum.

LC-MS: Rt=0.48 min; MS m/z [M+H]⁺ 661.5/663.4, m/z [M−H]⁻ 659.5/661.4;UPLC-MS 1

Intermediate T: tert-butyl4-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylateStep 1:3-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-1H-1,2,4-triazol-5-amine

6-Oxa-3-azabicyclo[3.1.1]heptane.HCl (1.25 g, 9.22 mmol) was dissolvedin ACN (16 mL) and Et₃N (1.34 mL, 9.68 mmol). Dimethylcyanocarbonimidodithioate (1.50 g, 9.22 mmol) was added and the RM wasstirred at reflux for 19.75 hours. 6-Oxa-3-azabicyclo[3.1.1]heptane.HCl(160 mg, 1.18 mmol) dissolved in ACN (500 μL) and Et₃N (170 μL, 1.23mmol) was added and the RM was stirred at reflux for 20 hours, then itwas cooled to 0° C. Hydrazine hydrate (508 mg, 10.1 mmol) was addeddropwise to the thick suspension within 1 minute. The suspension wasstirred at reflux for 7 hours. Hydrazine hydrate (320 mg, 6.36 mmol) wasadded again and the RM was continued stirring at reflux for 20 hours.The RM was cooled to RT and concentrated under reduced pressure. Theresidue was taken up in EtOAc and concentrated to give the titlecompound as a beige solid.

LC-MS: Rt=0.25 min; MS m/z [M+H]⁺ 182.1; UPLC-MS 4

Step 2: tert-butyl4-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

3-(6-Oxa-3-azabicyclo[3.1.1]heptan-3-yl)-1H-1,2,4-triazol-5-amine (1.25g, 4.35 mmol) and tert-butyl4-(1-methoxy-1,3-dioxopentan-2-yl)piperazine-1-carboxylate (IntermediateEH) (1.64 g, 5.22 mmol) were dissolved in EtOH (12 mL) and H₃PO₄ (526mg, 4.56 mmol) was added. The RM was stirred at 90° C. for 6.5 hours,then at RT overnight. Tert-butyl4-(1-methoxy-1,3-dioxopentan-2-yl)piperazine-1-carboxylate (IntermediateEH) (350 mg, 1.11 mmol) was added again and the RM was stirred at 90° C.for 4 hours. The red solution was cooled to RT and DIPEA (2.28 mL, 13.0mmol) was added, followed by Boc₂O (426 mL, 1.83 mmol). The RM wasstirred at RT for 1 hour and was quenched with water. The solvent wasremoved under reduced pressure and the aqueous layer was extracted withEtOAc (3×60 mL), aq sat NaHCO₃ (2×20 mL) and brine (2×20 mL). Thecombined organic layers were dried through a phase separator andconcentrated under reduced pressure. The residue was adsorbed ontoIsolute and purified by column chromatography (RediSep Column: Silica 40g, eluent DCM:MeOH 100:0 to 85:15). The product containing fractionswere combined, concentrated and dried under HV to give the titlecompound as an orange solid.

LC-MS: Rt=0.94 min; MS m/z [M+H]⁺ 446.2, m/z [M−H]⁻ 444.2; UPLC-MS 3

Intermediate U:2-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate T) (316 mg, 709 μmol) and2-iodo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (Intermediate DV)(256 mg, 745 μmol) were mixed with DMF (2.7 mL) at 0° C. DIPEA (372 μL,2.13 mmol) was added slowly. The RM was stirred at RT for 3 hours, thenit was stored in the freezer overnight. The next day it was continuedstirring at RT for 5 hours. The RM was quenched with water (5 mL) andstirred at RT. The suspension was filtered and washed with water to givethe title compound as a bright brown solid.

LC-MS: Rt=1.17 min; MS m/z [M+H]⁺ 661.3, m/z [M−H]⁻ 659.2; UPLC-MS 3

Step 2:2-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Tert-butyl4-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(426 mg, 645 μmol) was dissolved in DCM (4.4 mL) and TFA (745 μL, 9.67mmol) was added. The RM was stirred at RT for 25 minutes, then it wascooled with dry ice and Et₃N (1.79 mL, 12.9 mmol) was added. The RM wasconcentrated under reduced pressure. The residue was adsorbed ontoIsolute and purified by column chromatography (RediSep Column: Silica 40g, eluent DCM:MeOH 100:0 to 75:25). The product containing fractionswere combined, concentrated under reduced pressure and dried under HV togive the title compound as a bright brown solid.

LC-MS: Rt=0.81 min; MS m/z [M+H]⁺ 561.5, m/z [M−H]⁻ 559.3; UPLC-MS 3

Intermediate V:2-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-5-methyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

3-(6-Oxa-3-azabicyclo[3.1.1]heptan-3-yl)-1H-1,2,4-triazol-5-amine(Intermediate T step 1) (1.25 g, 4.35 mmol) and tert-butyl4-(1-ethoxy-1,3-dioxobutan-2-yl)piperazine-1-carboxylate (IntermediateEP) (1.64 g, 5.22 mmol) were mixed in EtOH (12 mL). H₃PO₄ 85% (526 mg,4.56 mmol) was added and the RM was stirred at 90° C. for 3 hours, thenit was cooled to RT. The RM was diluted with water and the resultingsuspension was filtered. The cake was discarded. The filtrate wasconcentrated under reduced pressure. The residue was extracted threetimes with EtOAc (3×80 mL), once with aq sat NaHCO₃ (25 mL) and oncewith brine (25 mL). The combined organic layers were dried through aphase separator and concentrated under reduced pressure to give crudeproduct. The aqueous layer still contained product so it wasconcentrated under reduced pressure. The solid residue was suspended inEtOAc and filtered. The cake still contained product so it was suspendedin DCM/MeOH and filtered. The filtrates were combined and concentratedunder reduced pressure. The residue was combined with the crude productto give the title compound as a brown solid foam.

LC-MS: Rt=0.87 min; MS m/z [M+H]⁺ 432.4, m/z [M−H]⁻ 430.4; UPLC-MS 3

Step 2: tert-butyl4-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-4-(2-((2-fluoro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(399 mg, 786 μmol) and2-bromo-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide (IntermediateDR) (259 mg, 865 μmol) were mixed in DMF (3 mL) at RT. DIPEA (412 μL,2.36 mmol) was added slowly. The suspension was stirred at RT for 1.5hours. Water (10 mL) was added and the mixture was continued stirring atRT. The suspension was filtered and washed with water. The crude productwas adsorbed onto Isolute and purified by column chromatography (RediSepColumn: Silica 40 g, eluent DCM:MeOH 100:0 to 90:10). The productcontaining fractions were combined and concentrated to give the titlecompound as a pale beige solid.

LC-MS: Rt=1.12 min; MS m/z [M+H]⁺ 651.4, m/z [M−H]⁻ 649.4; UPLC-MS 3

Step 3:2-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-5-methyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide

Tert-butyl4-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-4-(2-((2-fluoro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(353 mg, 543 μmol) was dissolved in DCM (3.7 mL) and TFA (836 μL, 10.9mmol) was added. The RM was stirred at RT for 1 hour. The RM wasconcentrated under reduced pressure, dissolved and concentrated twice inDCM and toluene. Then it was dried under HV to give the title compoundas a beige solid foam.

LC-MS: Rt=0.79 min; MS m/z [M+H]⁺ 551.3, m/z [M−H]⁻ 549.3; UPLC-MS 3

Intermediate W:2-(2-((3R,4S)-3,4-difluoropyrrolidin-1-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: 3-((3R,4S)-3,4-difluoropyrrolidin-1-yl)-1H-1,2,4-triazol-5-amine

(3S,4R)-3,4-difluoropyrrolidine.HCl (1.50 g, 10.5 mmol) was dissolved inACN (18 mL) and Et₃N (1.52 mL, 11.0 mmol). Dimethylcyanocarbonimidodithioate (2.01 g, 12.9 mmol) was added and the RM wasstirred at reflux for 2.5 hours. (3S,4R)-3,4-difluoropyrrolidine.HCl(310 mg, 2.16 mmol) and Et₃N (170 μL, 1.23 mmol) were added again andthe RM was stirred at reflux for 18.5 hours. The RM was cooled to 0° C.and hydrazine hydrate (710 mg, 14.2 mmol) was added within 1 minute tothe suspension. The RM was stirred at reflux for 23.5 hours. Thesuspension turned into a solution. Hydrazine hydrate (500 mg, 10.0 mmol)was added and the RM was stirred at 88° C. for 3 hours. The RM wascooled to 0° C. and the suspension was filtered and washed with coldEtOH. The obtained solid was dried under HV to give the title compoundas a beige solid.

LC-MS: Rt=0.29 min; MS m/z [M+H]⁺ 190.2, m/z [M−H]⁻ 188.1; UPLC-MS 3

Step 2: tert-butyl4-(2-((3R,4S)-3,4-difluoropyrrolidin-1-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

3-((3R,4S)-3,4-difluoropyrrolidin-1-yl)-1H-1,2,4-triazol-5-amine (1.25g, 6.61 mmol) and tert-butyl4-(1-methoxy-1,3-dioxopentan-2-yl)piperazine-1-carboxylate (IntermediateEH) (2.29 g, 7.27 mmol) were dissolved in EtOH (10 mL) and H₃PO₄ (85%)(800 mg, 6.94 mmol) was added slowly. Additional EtOH (10 mL) was addedand RM was stirred at 90° C. for 21 hours. Tert-butyl4-(1-methoxy-1,3-dioxopentan-2-yl)piperazine-1-carboxylate (IntermediateEH) (700 mg, 2.22 mmol) was added and the mixture was continued stirringat 90° C. for 24 hours. Tert-butyl4-(1-methoxy-1,3-dioxopentan-2-yl)piperazine-1-carboxylate (IntermediateEH) (500 mg, 1.59 mmol) was added and the RM was continued stirring at90° C. for 7 hours. The RM was allowed to cool to RT then DIPEA (3.46mL, 19.8 mmol) and Boc₂O (600 mg, 2.75 mmol) were added and the RM wasstirred at RT for 2 hours. Then water was added and the organic solventwas evaporated under reduced pressure. The aqueous layer was extractedwith EtOAc (3×60 mL), aq sat NaHCO₃ (2×20 mL) and brine (2×20 mL). Thecombined organic layers were dried through a phase separator andconcentrated under reduced pressure. The aqueous layer still containedproduct so it was concentrated under reduced pressure. The residue wassuspended several times in DCM and a small amount of MeOH and filtered.The residue was combined with the concentrated organic layer. It wasdissolved in DCM and concentrated to ⅓ of the volume. Then it wasstanding around at RT overnight, the resulting suspension was filteredand washed with a small amount of DCM to obtain product as a solid. Thefiltrate was adsorbed onto Isolute and purified by column chromatography(RediSep Column: Silica 40 g, eluent DCM:MeOH 100:0 to 90:10). Theproduct containing fractions were combined, concentrated, combined withthe cake and dried under HV to give the title compound as a yellowsolid.

LC-MS: Rt=0.95 min; MS m/z [M+H]⁺ 454.4, m/z [M−H]⁻ 452.4; UPLC-MS 3

Step 3: tert-butyl4-(2-((3R,4S)-3,4-difluoropyrrolidin-1-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-((3R,4S)-3,4-difluoropyrrolidin-1-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(600 mg, 1.32 mmol) and2-iodo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (Intermediate DV)(499 mg, 1.46 mmol) were dissolved in DMF (6 mL) and DIPEA (693 μL, 3.97mmol) was added. The RM was stirred at 30° C. for 2 hours, then at 40°C. for 4 hours. 2-lodo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate DV) (35.0 mg, 102 μmol) was added again and the RM wasstirred at 40° C. for 2.75 hours, then it was cooled to RT overnight.2-lodo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (Intermediate DV)(60.0 mg, 176 μmol) was added again and the RM was stirred at 40° C. for1.5 hours. The RM was quenched with water and stirred at RT. Theresulting suspension was filtered off and washed with a small amount ofwater. The crude product was adsorbed onto Isolute and purified bycolumn chromatography (RediSep Column: Silica 40 g, eluent DCM:MeOH100:0 to 90:10). The product containing fractions were combined,concentrated and dried under HV to give the title compound as a palebeige solid.

LC-MS: Rt=1.20 min; MS m/z [M+H]⁺ 669.4, m/z [M−H]⁻ 667.4; UPLC-MS 3

Step 4:2-(2-((3R,4S)-3,4-difluoropyrrolidin-1-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Tert-butyl4-(2-((3R,4S)-3,4-difluoropyrrolidin-1-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(410 mg, 613 μmol) was dissolved in DCM (4.1 mL) and TFA (709 μL, 9.20mmol) was added and the RM was stirred at RT for 2 hours. The RM wasconcentrated under reduced pressure. The residue was dissolved threetimes in DCM and a small amount of toluene and concentrated underreduced pressure again. Then it was dried under HV to give the titlecompound as a bright brown solid.

LC-MS: Rt=0.85 min; MS m/z [M+H]⁺ 569.4, m/z [M−H]⁻ 567.3; UPLC-MS 3

Intermediate X: tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

3-Bromo-1H-1,2,4-triazol-5-amine (Intermediate ER) (82.6 g, 507 mmol)and tert-butyl4-(1-methoxy-1,3-dioxopentan-2-yl)piperazine-1-carboxylate (IntermediateEH) (175 g, 557 mmol) were mixed in EtOH (465 mL). H₃PO₄ (49.7 g, 507mmol) was added. The mixture was stirred at 80° C. for 12 hours undernitrogen. The mixture was concentrated in vacuo to remove EtOH, thenquenched by addition of aq sat NaHCO₃ (1 L), and extracted with DCM (3×1L). The combined organic layers were washed with brine (3×1 L), driedover Na₂SO₄, filtered and concentrated under reduced pressure to give aresidue. The residue was purified by column chromatography (Silicacolumn, eluent DCM:MeOH 1:0 to 10:1). The product containing fractionswere combined and concentrated under reduced pressure to give the titlecompound as a yellow solid.

LC-MS: Rt=0.91 min; MS m/z [M+H−Boc]⁺ 327.1/329.1, m/z [M+H]⁺427.2/429.2, m/z [M−H]⁻ 425.2/427.2; UPLC-MS 1

LC-MS: Rt=4.53 min; MS m/z [M+H−Boc]⁺ 327.1/329.1, m/z [M−H]⁻425.2/427.2; UPLC-MS 2

¹H NMR (400 MHz, DMSO-d₆) δ 13.27 (s, 1H), 3.91 (m, 2H), 3.31 (m, 2H),2.88 (m, 2H), 2.75 (m, 2H), 2.61 (m, 2H), 1.42 (s, 9H), 1.17 (t, J=7.4Hz, 3H)

Intermediate Y:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (1.40 g, 3.28 mmol) was suspended in DMF/Water (9:1)(20 mL).Rac-2-(4-methoxycyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(Intermediate DH) (1.56 g, 6.55 mmol) and K₃PO₄ (2.09 g, 9.83 mmol) wereadded and the RM was degassed for 15 minutes with argon, thenPd(dppf)Cl₂.DCM (134 mg, 164 μmol) was added and the RM was stirred at100° C. for 16 hours. The RM was extracted three times with DCM. Theorganic layer was washed with brine, dried over Na₂SO₄ and concentratedunder reduced pressure. The crude product was purified by columnchromatography (Silica gel column: Silica 40 g, eluent DCM:MeOH 100:0 to98:2). The product containing fractions were combined and concentratedunder reduced pressure to give the title compound.

LC-MS: Rt=2.15 min; MS m/z [M+H]+ 459.3; UPLC-MS 13

Step 2: tert-butyl4-(4-(2-((2-chloro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(1.20 g, 2.54 mmol) and2-bromo-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide(Intermediate DQ) (645 mg, 2.03 mmol) were suspended in DMF (8 mL) at 0°C. and DIPEA (1.33 mL, 7.62 mmol) was added dropwise. The RM was stirredat RT for 14 hours. The RM was concentrated, extracted with 10% MeOH inDCM, dried over Na₂SO₄ and concentrated under reduced pressure. Thecrude product was purified by column chromatography (Silica gel column:Silica 12 g, eluent DCM:MeOH 100:0 to 98:2). The product containingfractions were combined and concentrated to give the title compound.

LC-MS: Rt=2.41 min; MS m/z [M+H]⁺ 695.7/697.7; UPLC-MS 13

Step 3:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((2-chloro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(400 mg, 529 μmol) was suspended in DCM (15 mL) and TFA (8.00 mL, 104mmol) was added at 0° C. The RM was stirred at RT for 16 hours. The RMwas quenched with NaHCO₃ and extracted with DCM. The organic layer wasdried over Na₂SO₄ and concentrated under reduced pressure. The solid wastriturated with Et₂O to give the title compound.

LC-MS: Rt=0.51 min; MS m/z [M+H]⁺ 595.1/597.1; UPLC-MS 12

Intermediate Z:rac-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamideStep 1: rac-tert-butyl4-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)iperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (1.00 g, 2.34 mmol) was suspended in DMF/Water (9:1)(15 mL).Rac-2-(4-methoxycyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(Intermediate DH) (1.12 g, 4.68 mmol) and K₃PO₄ (1.49 g, 7.02 mmol) wereadded and the RM was degassed for 15 minutes with argon, then XPhos PdG2 (92.0 mg, 117 μmol) was added and the RM was stirred at 110° C. for12 hours. The RM was concentrated, ice cold water was added and theprecipitate was filtered off, washed with Et₂O and dried under reducedpressure. The crude product was purified by column chromatography(Silica gel column: Silica 40 g, eluent DCM:MeOH 100:0 to 97:3). Theproduct containing fractions were combined and concentrated underreduced pressure to give the title compound.

LC-MS: Rt=1.48 min; MS m/z [M+H]⁺ 459.2; UPLC-MS 11

Step 2: rac-tert-butyl4-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-4-(2-((2-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To the solution of rac-tert-butyl4-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(400 mg, 872 μmol) in DMF (8 mL) were added2-bromo-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide(Intermediate DW) (311 mg, 1.05 mmol) and DIPEA (457 μL, 2.62 mmol) andthe RM was stirred at RT for 16 hours. The RM was diluted with water andextracted with 5% MeOH in DCM, washed with brine, dried over Na₂SO₄ andconcentrated. The crude product was purified by column chromatography(Silica gel column: Silica 24 g, eluent DCM:MeOH 100:0 to 98:2). Theproduct containing fractions were combined, concentrated and dried underHV to give the title compound.

LC-MS: Rt=2.33 min; MS m/z [M+H]⁺ 675.2; UPLC-MS 13

Step 3:rac-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

To the solution of rac-tert-butyl4-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-4-(2-((2-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(400 mg, 414 μmol) in DCM (5 mL) was added 1,4-dioxane 4M in HCl (10.0mL, 40.0 mmol) at 0° C. and the RM was stirred at RT for 12 hours. TheRM was concentrated under reduced pressure, washed with Et₂O (2×10 mL)and dried under HV to give the title compound as a HCl salt as anoff-white solid.

LC-MS: Rt=1.31 min; MS m/z [M+H]⁺ 575.3; UPLC-MS 11

Intermediate AA:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (600 mg, 1.40 mmol),2-(5,6-dihydro-1,4-dioxin-2-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(357 mg, 1.69 mmol) and XPhos Pd G3 (59.4 mg, 70.0 μmol) were mixed,evaporated and purged with argon several times. 1,4-Dioxane (10 mL) andK₃PO₄ 1M in water (4.21 mL, 4.21 mmol) were added and the RM was stirredat 80° C. for 4.75 hours.2-(5,6-dihydro-1,4-dioxin-2-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(90.0 mg, 425 μmol) was added and the mixture was continued stirring at80° C. for 1.5 hours.2-(5,6-dihydro-1,4-dioxin-2-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(110 mg, 519 μmol) and XPhos Pd G3 (15.0 mg, 17.7 μmol) were added andthe RM was stirred at 80° C. for 45 minutes. Then it was stirred at RTovernight.2-(5,6-dihydro-1,4-dioxin-2-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(60.0 mg, 283 μmol) was added and the RM was stirred at 80° C. for 2.5hours. The RM was cooled to RT and water (5 mL) and aq sat NaHCO₃ (5 mL)were added. The RM was extracted with EtOAc (3×40 mL). The organic layerwas washed with water (10 mL), dried through a phase separator andconcentrated under reduced pressure. The aqueous layer was washed withDCM (4×40 mL) and with brine, then 4 times with 10% MeOH in DCM. Theorganic layer was dried through a phase separator and concentrated underreduced pressure. The organic layers were combined and ISOLUTE® Si-Thiol(1.50 g) was added. The mixture was stirred at 35° C. for 30 minutes.The suspension was filtered and washed with DCM and MeOH, the filtratewas concentrated under reduced pressure. The crude product was dissolvedin DCM (15 mL) and sonicated for 2 minutes. The suspension was filtered,washed with a small amount of DCM and dried to give the title compoundas a white solid.

LC-MS: Rt=0.88 min; MS m/z [M+H]⁺ 433.4, m/z [M−H]⁻ 431.4; UPLC-MS 1

Step 2: tert-butyl4-(4-(2-((2-chloro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(756 mg, 1.40 mmol) andN-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-iodoacetamide(Intermediate DU) (510 mg, 1.40 mmol) were mixed with DMF (6.1 mL) atRT. DIPEA (733 μL, 4.20 mmol) was added and the RM was stirred at RT for2.75 hours. The RM was stored in the fridge overnight. The next morningit was stirred at RT for 21 hours. The RM was quenched with water (25mL) and the mixture was stirred at RT. The suspension was filtered,washed with water and the cake was dried. The filtrate was extractedwith DCM (4×40 mL). The organic layer was dried through a phaseseparator and concentrated under reduced pressure. The resulting solidwas combined with the cake. The crude product was adsorbed onto Isoluteand purified by column chromatography (RediSep Column: Silica 24 g,eluent DCM:MeOH 100:0 to 85:15). The product containing fractions werecombined, concentrated and dried under HV to give the title compound asa beige solid.

LC-MS: Rt=1.18 min; MS m/z [M+H−Boc]⁺ 569.3/571.3, m/z [M−H]⁻667.2/669.1; UPLC-MS 1

Step 3:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((2-chloro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(462 mg, 380 μmol) was dissolved in DCM (5 mL). TFA (798 μL, 10.4 mmol)was added and the RM was stirred at RT for 1.5 hours. Then it wasconcentrated under reduced pressure. The residue was dissolved in DCMand a small amount of toluene and concentrated under reduced pressure.This was performed three times. The residue was dried under HV to givethe title compound as a beige solid.

LC-MS: Rt=0.64 min; MS m/z [M+H]⁺ 569.3/571.3, m/z [M−H]⁻ 567.1/569.1;UPLC-MS 1

Intermediate AB: tert-butyl4-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (5.00 g, 11.7 mmol) was suspended in DMF/water (9:1)(50 mL).2-(3,4-dihydro-2H-pyran-6-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(3.69 g, 17.6 mmol) and K₃PO₄ (7.45 g, 35.1 mmol) were added and the RMwas degassed with argon for 15 minutes. Pd(dppf)Cl₂.DCM (478 mg, 585μmol) was added and the RM was stirred at 100° C. for 12 hours. The RMwas diluted with DCM and washed with water. The organic layer was driedover Na₂SO₄ and concentrated under reduced pressure. The insoluble solidwas filtered and washed with n-hexane and Et₂O and dried. The solid wasdissolved in 10% MeOH in DCM and washed with water and brine. Theorganic layer was concentrated under reduced pressure. The crude productwas purified twice by column chromatography (2× Silica gel column:Silica 40 g, eluent DCM:MeOH 100:0 to 95:5). The product containingfractions were combined, concentrated under reduced pressure and driedto give the title compound.

LC-MS: Rt=2.12 min; MS m/z [M+H]⁺ 431.1; UPLC-MS 13

Intermediate AC:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((2-chloro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To the stirred solution of tert-butyl4-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate AB) (630 mg, 1.46 mmol) in DMF (10 mL) were added2-bromo-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide(Intermediate DQ) (441 mg, 1.39 mmol) and DIPEA (1.02 mL, 5.85 mmol) at0° C. The RM was stirred at RT for 16 hours. The RM was concentrated,ice cold water was added, the precipitate was filtered off and driedunder HV. The crude product was purified by column chromatography(Silica gel column: Silica 12 g, eluent DCM:MeOH 100:0 to 90:10). Theproduct containing fractions were combined and concentrated to give thetitle compound as a brown solid.

LC-MS: Rt=1.65 min; MS m/z [M+H]⁺ 667.2/669.2; UPLC-MS 11

Step 2:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((2-chloro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(650 mg, 818 μmol) was suspended in DCM (10 mL). Et₃N (567 μL, 4.09mmol) and TMSOTf (887 μL, 4.91 mmol) were added at RT and the RM wasstirred at 0° C. to RT for 4 hours. The RM was concentrated underreduced pressure to give the title compound.

LC-MS: Rt=1.34 min; MS m/z [M+H]⁺ 567.2/569.2; UPLC-MS 11

Intermediate AD:2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamideStep 1: tert-butyl4-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-4-(2-((2-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate AB) (2.00 g, 4.65 mmol) was dissolved in DMF (20 mL) and2-bromo-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide(Intermediate DW) (1.38 g, 4.65 mmol) and DIPEA (2.43 mL, 13.9 mmol)were added and the RM was stirred at RT for 16 hours. The RM was dilutedwith water, extracted with 5% MeOH in DCM, washed with brine, dried overNa₂SO₄ and concentrated under reduced pressure. The crude product waspurified by column chromatography (Silica gel column: Silica 40 g,eluent DCM:MeOH 100:0 to 98:2). The product containing fractions werecombined, concentrated and dried. The still impure product was purifiedagain by column chromatography (Silica gel column: Silica 40 g, eluentDCM:MeOH 100:0 to 98:2). The product containing fractions were combined,concentrated and dried to give the title compound.

LC-MS: Rt=1.6 min; MS m/z [M−H]⁻ 645.3; UPLC-MS 11

Step 2:2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

To the stirred solution of tert-butyl4-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-4-(2-((2-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(1.00 g, 1.44 mmol) in DCM (20 mL) at 0° C. were added Et₃N (1.20 mL,8.63 mmol) and TMSOTf (1.56 mL, 8.63 mol) and the RM was stirred at 0°C. for 4 hours, then at RT for 16 hours. The RM was concentrated and icecold water was added. The mixture was stirred for 30 minutes. Theprecipitate was filtered off, washed with water (5 mL) and n-pentane anddried under HV to give the title compound.

LC-MS: Rt=1.34 min; MS m/z [M+H]⁺ 547.3; UPLC-MS 11

Intermediate AE:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate AB) (5.05 g, 6.92 mmol) andN-(2-chloro-4-(trifluoromethyl)phenyl)-2-iodoacetamide (Intermediate DL)(1.85 g, 5.09 mmol) were mixed in 1,4-dioxane (20 mL) and DIPEA (2.42mL, 13.8 mmol) was added. The RM was stirred at 80° C. for 30 minutes.Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL) were added. Theaqueous layer was washed twice with DCM (2×10 mL). The combined organiclayers were dried through a phase separator and concentrated underreduced pressure. The crude product was purified by columnchromatography (RediSep Column: Silica 80 g, eluent DCM:DCM/MeOH (1/1)100:0 to 70:30). The product containing fractions were combined,concentrated under reduced pressure and dried under HV to afford a brownoil. It was purified by column chromatography again (RediSep Column:Silica 40 g, eluent DCM:DCM/MeOH (8/2) 100:0 to 45:55). The productcontaining fractions were combined, concentrated under vacuum and driedunder HV to afford the title compound as a brown oil.

LC-MS: Rt=1.36 min; MS m/z [M−H]⁻ 664.5/666.5; UPLC-MS 1

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(763 mg, 882 μmol) was dissolved in DCM (5 mL) and TFA (1.00 mL, 13.0mmol) was added. The mixture was stirred at 40° C. for 1 hour. Themixture was concentrated under reduced pressure. Water (10 mL), aq satNaHCO₃ (10 mL) and DCM (10 mL) were added. The aqueous layer was washedtwice with DCM (2×10 mL). The combined organic layers were dried througha phase separator and concentrated under reduced pressure. The crudeproduct was suspended in Et₂O and sonicated for 5 minutes. Then it wasfiltered. The cake was washed with Et₂O and dried under HV to give thetitle compound.

LC-MS: Rt=0.87 min; MS m/z [M+H]⁺ 566.3/568.3, m/z [M−H]⁻ 564.4/566.4;UPLC-MS 1

Intermediate AF: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To the stirred solution of tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (15.0 g, 35.1 mmol) in 1,4-dioxane (150 mL) and water(50 mL) was added2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(11.1 g, 52.7 mmol) and Na₂CO₃ (7.44 g, 70.2 mmol). The RM was degassedwith nitrogen for 15 minutes. Pd(dppf)Cl₂.DCM (1.43 g, 1.76 mmol) wasadded and the RM was stirred at 100° C. for 14 hours. Water (300 mL) wasadded and the RM was extracted with 10% MeOH in DCM (2×500 mL). Theorganic layer was washed with brine (300 mL), dried over Na₂SO₄ andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (Silica gel column: Silica 40 g, eluent DCM:MeOH100:0 to 97:3). The product containing fractions were combined,concentrated under vacuum and dried under HV to give the title compound.

LC-MS: Rt=0.96 min; MS m/z [M+H]⁺ 431.4, m/z [M−H]⁻ 429.3; UPLC-MS 3

¹H NMR (400 MHz, DMSO-d₆) δ 13.00 (s, br, 1H), 6.81 (m, 1H), 4.28 (m,2H), 3.92 (m, 2H), 3.82 (m, 2H), 3.37 (m, 2H), 2.89 (m, 2H), 2.76 (m,2H), 2.62 (m, 2H), 2.51 (m, 2H), 1.43 (s, 9H), 1.19 (t, J=7.3 Hz, 3H)

Intermediate AG:N-(4-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((4-chloro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate AF) (610 mg, 1.42 mmol) was suspended in DMF (5 mL).2-Chloro-N-(4-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide(Intermediate DX) (348 mg, 1.28 mmol) and DIPEA (619 μL, 3.54 mmol) wereadded at 0° C. and stirred at 70° C. for 24 hours. Water was added andthe solid was filtered. Then it was washed with Et₂O and dried. Thecrude product was purified by column chromatography (Silica gel column:Silica 24 g, eluent DCM:MeOH 100:0 to 99:1). The product containingfractions were combined, concentrated and dried to give the titlecompound.

LC-MS: Rt=2.33 min, MS m/z [M+H]⁺ 667.3/669.3; UPLC-MS 13

Step 2:N-(4-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((4-chloro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(500 mg, 750 μmol) was suspended in DCM (10 mL) and TFA (5.00 mL, 64.9mmol) was added at 0° C. The RM was stirred at RT for 12 hours. The RMwas concentrated under reduced pressure to give the title compound.

LC-MS: Rt=1.34 min, MS m/z [M+H]⁺ 567.2/569.2; UPLC-MS 11

Intermediate AH:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamideStep 1: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((4-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate AF) (400 mg, 929 μmol) was suspended in DMF (5 mL).2-Bromo-N-(4-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide(Intermediate DY) (248 mg, 836 μmol) and DIPEA (406 μL, 2.32 mmol) wereadded at 0° C. and the RM was stirred at RT for 12 hours. Water wasadded and the solid was filtered. Then it was washed with Et₂O and driedto give the title compound.

LC-MS: Rt=1.55 min, MS m/z [M+H−Boc]⁺ 547.2; UPLC-MS 11

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((4-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(450 mg, 70% pure, 487 μmol) was suspended in DCM (10 mL) and TFA (5.00mL, 64.9 mmol) was added at 0° C. The RM was stirred at RT for 12 hours.The RM was concentrated under reduced pressure to give the titlecompound.

LC-MS: Rt=0.72 min, MS m/z [M+H]⁺ 547.3; UPLC-MS 13

Intermediate AI:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamideStep 1: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To a solution of tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate AF) (3.00 g, 6.97 mmol) in DMF (30 mL) was added DIPEA(3.65 mL, 20.9 mmol) and2-bromo-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide(Intermediate DW) (1.66 g, 5.57 mmol) at 0° C., then allowed to warm toRT and stirred at RT for 18 hours. The RM was quenched with water andacidified with HCl. Then it was extracted with DCM, dried over Na₂SO₄and concentrated under reduced pressure. The crude product was purifiedby column chromatography (Silica gel column: Silica 40 g, eluenthexane:EtOAc 100:0 to 40:60). The product containing fractions werecombined, concentrated and washed with Et₂O to give the title compound.

LC-MS: Rt=1.59 min; MS m/z [M−H]⁻ 645.3; UPLC-MS 13

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(2.50 g, 3.75 mmol) was taken in DCM (25 mL) and TFA (8.00 mL, 104 mmol)was added. The RM was stirred at RT for 2 hours. The crude product wasconcentrated under reduced pressure, washed with Et₂O and dried under HVto give the title compound, as the TFA salt.

LC-MS: Rt=0.45 min, MS m/z [M+H]⁺ 547.6; UPLC-MS 13

Intermediate AJ:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate AF) (2.00 g, 4.55 mmol) andN-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)-2-iodoacetamide(Intermediate DZ) (2.26 g, 4.82 mmol) were mixed in 1,4-dioxane (20 mL)and DIPEA (1.59 mL, 9.11 mmol) was added. The mixture was stirred at 70°C. for 1.5 hours. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL)were added. The aqueous layer was washed twice with DCM (2×10 mL). Thecombined organic layers were dried through a phase separator andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (RediSep Column: Silica 80 g, eluent DCM:DCM/MeOH(8/2) 100:0 to 65:35). The product containing fractions were combined,concentrated under vacuum and dried under HV to afford the titlecompound.

LC-MS: Rt=1.30 min; MS m/z [M+H−Boc]⁺ 564.2, m/z [M−H]⁻ 662.5; UPLC-MS 1

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(2.18 g, 3.02 mmol) was dissolved in DCM (15 mL) and TFA (1.00 mL, 13.0mmol) was added. The RM was stirred at 40° C. for 3 hours. About 50%conversion was observed. The RM was concentrated under reduced pressureto store it over the weekend, then it was restarted with DCM (15 mL) andTFA (1.00 mL, 13.0 mmol). The RM was stirred at 40° C. for 3 hours.Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL) were added. Theaqueous layer was washed twice with DCM (2×10 mL). The combined organiclayers were dried through a phase separator and concentrated underreduced pressure to give the title compound.

LC-MS: Rt=0.81 min; MS m/z [M+H]⁺ 564.2, m/z [M−H]⁻ 562.4; UPLC-MS 1

Intermediate AK:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate AF) (6.50 g, 15.1 mmol) andN-(2-chloro-4-(trifluoromethyl)phenyl)-2-iodoacetamide (Intermediate DL)(6.04 g, 16.6 mmol) were mixed in DMF (72 mL) at 0° C. DIPEA (7.91 mL,45.3 mmol) was added and the RM was stirred at 45° C. for 3.5 hours. TheRM was cooled to RT. Water (70 mL) was added and the suspension wasstirred at RT overnight. The suspension was sonicated for 25 minutes andfiltered. The cake was washed with a small amount of water and dried.The filtrate was filtered again. The second filtrate was extracted withEtOAc (2×400 mL), washed with brine (2×50 mL), dried through a phaseseparator and concentrated under reduced pressure. The 2 cakes wereadsorbed onto Isolute and purified by column chromatography (RediSepColumn: Silica 220 g, eluent DCM:DCM/MeOH (1/1) 100:0 to 80:20). Thepure product containing fractions were combined and concentrated underreduced pressure. The beige solid foam was dissolved in Et₂O and theresulting crystals were sonicated. The suspension was left standingovernight, filtered, washed with a small amount of Et₂O and dried underHV to give cake 1 as a white solid. The impure fractions were combinedand concentrated under reduced pressure. Then they were combined withthe concentrated organic layer from the extraction and purified bycolumn chromatography again (RediSep Column: Silica 120 g Gold, eluentDCM:DCM/MeOH (1/1) 100:0 to 85:15). The product containing fractionswere combined, concentrated under reduced pressure and dried under HV.The beige solid foam was crystallized out of Et₂O to give cake 2 as awhite solid. Cake 1 and cake 2 were combined to give the title compound.

LC-MS: Rt=1.33 min; MS m/z [M+H−Boc]⁺ 566.0/568.0, m/z [M+H]⁺666.0/668.0, m/z [M−H]⁻ 664.1/666.1; UPLC-MS 1

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(6.41 g, 9.62 mmol) was dissolved in DCM (70 mL) and TFA (11.1 mL, 144mmol) was added. The RM was stirred at RT for 1 hour. The RM wasconcentrated under reduced pressure. The residue was dissolved in DCMand concentrated under reduced pressure again. This was performed threetimes. The resulting oil was dried under HV to result in a pale rosesolid foam. The foam was suspended in Et₂O and sonicated. The suspensionwas filtered, washed with Et₂O and dried under HV to give the titlecompound as a white solid.

LC-MS: Rt=0.78 min; MS m/z [M+H]⁺ 566.4/568.4, m/z [M−H]⁻ 564.2/566.2;UPLC-MS 1

Intermediate AL:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((2-chloro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate AF) (344 mg, 719 μmol) andN-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-iodoacetamide(Intermediate DU) (365 mg, 791 μmol) were mixed in 1,4-dioxane (5 mL)and DIPEA (314 μL, 1.78 mmol) was added. The mixture was stirred at 80°C. for 4.5 hours. After stirring at RT for 2 days, it was stirred at 80°C. for 6 hours. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL)were added. The aqueous layer was washed twice with DCM (2×10 mL). Thecombined organic layers were dried through a phase separator andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (RediSep Column: Silica 24 g, eluent DCM:DCM/MeOH(9/1) 100:0 to 50:50). The product containing fractions were combined,concentrated under vacuum and dried under HV to afford the titlecompound as a brown solid.

LC-MS: Rt=1.14 min; MS m/z [M+H−Boc]⁺ 567.5/569.4, m/z [M−H]⁻665.3/667.3; UPLC-MS 3

Step 2:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((2-chloro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(342 mg, 513 μmol) was dissolved in DCM (5 mL) and TFA (1.00 mL, 13.0mmol) was added. The RM was stirred at 40° C. for 20 minutes. The RM wasconcentrated under reduced pressure. The crude product was purified in 2portions by reverse phase preparative HPLC (2×RP-HPLC acidic 1: 20 to55% B in 10 min with a plateau at 55% for 1 min). The product containingfractions were combined, basified with aq sat NaHCO₃, extracted twicewith DCM, dried through a phase separator and concentrated under reducedpressure to give the title compound.

LC-MS: Rt=0.78 min; MS m/z [M+H]⁺ 567.3/569.3, m/z [M−H]⁻ 565.1/567.1;UPLC-MS 3

Intermediate AM:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate AF) (800 mg, 1.86 mmol) was suspended in DMF (5 mL).2-Bromo-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate DN) (700 mg, 2.23 mmol) and DIPEA (974 μL, 5.57 mmol) wereadded at 0° C. and the RM was stirred at RT for 12 hours. Water wasadded and the solid was filtered and dried to give the title compound.

LC-MS: Rt=1.65 min; MS m/z [M+H]⁺ 664.2; UPLC-MS 11

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(800 mg, 1.00 mmol) was suspended in DCM (2 mL) and TFA (2.00 mL, 26.0mmol) was added at 0° C. The RM was stirred at RT for 12 hours. The RMwas concentrated under reduced pressure to give the title compound.

LC-MS: Rt=1.34 min, MS m/z [M+H]⁺ 564.2; UPLC-MS 11

Intermediate AN:N-(2-chloro-4-(pentafluorosulfanyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((2-chloro-4-(pentafluorosulfanyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate AF) (625 mg, 1.45 mmol) and2-chloro-N-(2-chloro-4-(pentafluorosulfanyl)phenyl)acetamide(Intermediate EA) (527 mg, 1.60 mmol) were mixed in 1,4-dioxane (10 mL)and DMF (5 mL). Then DIPEA (634 μL, 3.63 mmol) was added and the RM wasstirred at 80° C. for 3 days. Then it was stirred at 110° C. for 3 days.Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL) were added. Theaqueous layer was washed twice with DCM (2×10 mL). The combined organiclayers were dried through a phase separator and concentrated underreduced pressure. The crude product was purified by columnchromatography (RediSep Column: Silica 40 g, eluent DCM:DCM/MeOH (1/1)100:0 to 65:35). The product containing fractions were combined,concentrated under reduced pressure and dried under HV. The resultingimpure product was purified in 5 portions by reverse phase preparativeHPLC (5×RP-HPLC acidic 1: 5 to 95% B in 20 min with a plateau at 95% for1 min). The product containing fractions were combined, basified with aqsat NaHCO₃, extracted twice with DCM, dried through a phase separatorand concentrated under reduced pressure to give the title compound.

LC-MS: Rt=1.39 min; MS m/z [M+H−Boc]⁺ 624.4/626.4, m/z [M−H]⁻722.5/724.5; UPLC-MS 1

Step 2:N-(2-chloro-4-(pentafluorosulfanyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((2-chloro-4-(pentafluorosulfanyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(600 mg, 81% pure, 671 μmol) was dissolved in DCM (10 mL) and TFA (2.00mL, 26.0 mmol) was added. The RM was stirred at 40° C. for 1 hour. TheRM was concentrated under reduced pressure. Water (10 mL), aq sat NaHCO₃(10 mL) and DCM (10 mL) were added. The aqueous layer was washed twicewith DCM (2×10 mL). The combined organic layers were dried through aphase separator and concentrated under reduced pressure to give thetitle compound.

LC-MS: Rt=0.90 min; MS m/z [M+H]⁺ 624.3/626.2, m/z [M−H]⁻ 622.4/624.3;UPLC-MS 1

Intermediate AO:N-(4-(difluoromethyl)-5-fluoro-2-methylphenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((5-fluoro-4-formyl-2-methylphenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To the stirred solution of tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate AF) (3.00 g, 6.97 mmol) in DMF (30 mL) at 0° C. were added2-bromo-N-(5-fluoro-4-formyl-2-methylphenyl)acetamide (Intermediate EB)(1.91 g, 6.97 mmol) and DIPEA (3.65 mL, 20.9 mmol) and the RM wasstirred at RT for 12 hours. Water (50 mL) was added and the RM wasextracted with DCM (2×50 mL). The organic layer was washed with brine(50 mL), dried over Na₂SO₄ and concentrated under reduced pressure. Thecrude product was purified by column chromatography (Silica gel column:Silica 12 g, eluent DCM:MeOH 100:0 to 98:2). The product containingfractions were combined and concentrated to give the title compound.

LC-MS: Rt=1.73 min, MS m/z [M+H−Boc]⁺ 524.0; UPLC-MS 12

Step 2: tert-butyl4-(4-(2-((4-(difluoromethyl)-5-fluoro-2-methylphenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((5-fluoro-4-formyl-2-methylphenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(1.34 g, 2.15 mmol) in DCM (20 mL) at −78° C. was added DAST (1.14 mL,8.59 mmol). The RM was stirred at −78° C. for 30 minutes. Then removedfrom cooling bath and allowed to warm for 40 minutes. Continued stirringat RT for 1.3 hours. The RM was cooled to 0° C. and carefully quenchedby addition of aq sat NaHCO₃ (30 mL). Stirred the biphasic mixture at RTfor 20 minutes. The organic layer was separated by filtration through aphase separator and evaporated in vacuo to give a brown oil. The crudeproduct was purified by column chromatography (RediSep Column: Silica 40g, eluent DCM:MeOH 100:0 to 50:50). The product containing fractionswere combined and concentrated to give the title compound as a palebrown solid—contains aldehyde as major side-product

LC-MS: Rt=1.19 min; MS m/z [M+H−Boc]⁺ 546.5, m/z [M−H]⁻ 644.3; UPLC-MS 1

Step 3:N-(4-(difluoromethyl)-5-fluoro-2-methylphenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To tert-butyl4-(4-(2-((4-(difluoromethyl)-5-fluoro-2-methylphenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(1.25 g, 1.94 mmol) in DCM (5 mL) was added TFA (750 μL, 9.73 mmol) andthe RM was stirred at RT for 1 hour. The RM was evaporated in vacuo. Theresidue was partitioned between DCM (50 mL) and aq sat NaHCO₃ (20 mL).The organic layer was filtered through a phase separator and evaporatedin vacuo to give the title compound as a brown solid—contains aldehydeside-product.

LC-MS: Rt=0.64 min; MS m/z [M+H]⁺ 546.3, m/z [M−H]⁻ 544.2; UPLC-MS 1

Intermediate AD:2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamideStep 1: tert-butyl4-(2-bromo-5-ethyl-4-(2-((2-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (355 mg, 831 μmol) and2-iodo-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide(Intermediate EC) (370 mg, 914 μmol) were mixed in DMF (3.2 mL). DIPEA(435 μL, 2.49 mmol) was added and the RM was stirred at 30° C. for 2hours, then at 40° C. for 4.75 hours, then at RT overnight. Water (5 mL)was added and stirring was continued at RT for 1 hour. The suspensionwas filtered and washed with a small amount of water to give the titlecompound as a redbrown solid foam.

LC-MS: Rt=1.11 min; MS m/z [M+H−Boc]⁺ 543.2, m/z [M−H]⁻ 641.3; UPLC-MS 3

Step 2: tert-butyl4-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-4-(2-((2-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-4-(2-((2-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(413 mg, 642 μmol),2-(3,4-dihydro-2H-pyran-6-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(202 mg, 963 μmol) and XPhos Pd G3 (27.7 mg, 32.0 μmol) were degassedand purged with argon several times. 1,4-Dioxane (3 mL) was added,followed by K₃PO₄ 1M in water (1.93 mL, 1.93 mmol) and the RM wasstirred at 90° C. for 1.3 hours. The RM was concentrated under reducedpressure. The aqueous residue was extracted with EtOAc (3×50 mL), andthe organic phases were washed with water (2×10 mL) and brine (10 mL).The organic layer was dried through a phase separator. ISOLUTE® Si-Thiol(550 mg) was added and the mixture was stirred for 20 minutes. Thesuspension was filtered and the filtrate was concentrated under reducedpressure. The crude product was adsorbed onto Isolute and purified bycolumn chromatography (RediSep Column: Silica 40 g, eluent DCM:MeOH100:0 to 90:10). The product containing fractions were combined andconcentrated under reduced pressure to give the title compound as agrey-brown solid.

LC-MS: Rt=1.11 min; MS m/z [M+H−Boc]⁺ 547.3, m/z [M−H]⁻ 645.5; UPLC-MS 3

Step 3:2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

Tert-butyl4-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-4-(2-((2-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(414 mg, 608 μmol) was dissolved in DCM (4 mL) and TFA (937 μL, 12.2mmol) was added and the RM was stirred at RT for 1.3 hours. The RM wasconcentrated under reduced pressure. Then it was dissolved in DCM andconcentrated under reduced pressure again. This was performed threetimes. The resulting material was dried under HV to give the titlecompound as a brown solid.

LC-MS: Rt=0.83 min; MS m/z [M+H]⁺ 547.4, m/z [M−H]⁻ 545.4; UPLC-MS 3

Intermediate AP:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-bromo-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To a stirred solution of tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (15.0 g, 33.3 mmol) and2-iodo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (Intermediate DV)(13.2 g, 38.4 mmol) in 1,4-dioxane (200 mL) was added DIPEA (17.5 mL,100 mmol) at RT and the RM was stirred at 80° C. for 15 minutes. The RMwas concentrated under reduced pressure. The crude product was dilutedwith EtOAc and water, extracted once with EtOAc and the organic layerwas washed with brine, dried over Na₂SO₄ and concentrated under reducedpressure. The crude product was purified by column chromatography(eluent heptane:EtOAc 70:30 to 0:100). The product containing fractionswere combined, concentrated and triturated in Et₂O to give the titlecompound as a white solid.

LC-MS: Rt=1.31 min; MS m/z [M+H−Boc]⁺ 542.2, m/z [M−H]⁻ 640.5; UPLC-MS 1

Step 2: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To a stirred solution of tert-butyl4-(2-bromo-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(11.2 g, 17.4 mmol), K₃PO₄ (11.1 g, 52.2 mmol) and XPhos Pd G3 (736 mg,870 μmol) in 1,4-dioxane (100 mL) and water (50 mL) was added dropwiseat 80° C. a solution of2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(4.75 g, 22.6 mmol) in 1,4-dioxane (50 mL) and the RM was stirred at 80°C. for 30 minutes. The RM was diluted with DCM and water, extractedtwice with DCM and the combined organic extracts were washed with brine,dried over Na₂SO₄, concentrated under reduced pressure and triturated inEt₂O to give the title compound as a white solid.

LC-MS: Rt=1.26 min; MS m/z [M+H−Boc]⁺ 546.4, m/z [M−H]⁻ 644.3; UPLC-MS 1

Step 3:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

HCl 4N in 1,4-dioxane (30.0 mL, 120 mmoL) was added to tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(9.83 g, 15.2 mmol) and the RM was stirred at 0° C. for 2 hours. The RMwas diluted with DCM and NaHCO₃, extracted three times with 10% MeOH inDCM and the combined organic layers were dried over Na₂SO₄, concentratedunder reduced pressure and dried to give the title compound as a whitesolid.

LC-MS: Rt=0.75 min; MS m/z [M+H]⁺ 546.3, m/z [M−H]⁻ 544.4; UPLC-MS 1

Intermediate AQ:N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(2-bromo-4-(2-((5-chloro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (300 mg, 702 μmol) was suspended in DMF (3 mL) and2-bromo-N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate DM) (278 mg, 842 μmol) and DIPEA (368 μL, 2.11 mmol) wereadded at 0° C. The RM was stirred at RT for 12 hours. Water was addedand the precipitate was filtered off to give the title compound.

LC-MS: Rt=1.68 min; MS m/z [M+H]⁺ 676.1; UPLC-MS 11

Step 2: tert-butyl4-(4-(2-((5-chloro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-4-(2-((5-chloro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(400 mg, 479 μmol) was suspended in 1,4-dioxane (5 mL).2-(3,6-Dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(201 mg, 957 μmol) and 0.6M aq Na₂CO₃ (2 mL) were added and the RM wasdegassed for 10 minutes with argon. PdCl₂(PPh₃)₂ (33.6 mg, 48.0 μmol)was added and the RM was stirred at 90° C. for 12 hours. The RM wasconcentrated under reduced pressure, and the crude product was purifiedby column chromatography (Silica gel column: Silica 12 g, eluentDCM:MeOH 100:0 to 97:3). The product containing fractions were combined,concentrated and dried under HV to give the title compound.

LC-MS: Rt=1.66 min; MS m/z [M+H]⁺ 680.2; UPLC-MS 11

Step 3:N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((5-chloro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(200 mg, 232 μmol) was suspended in DCM (5 mL). TFA (500 μL, 6.49 mmol)was added and the RM was stirred at 40° C. for 3 hours. The RM wasconcentrated under reduced pressure to give the title compound.

LC-MS: Rt=1.37 min; MS m/z [M+H]⁺ 580.2; UPLC-MS 11

Intermediate AR:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamideStep 1: tert-butyl4-(2-bromo-5-ethyl-4-(2-((5-fluoro-2-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (663 mg, 1.55 mmol) andN-(5-fluoro-2-methyl-6-(trifluoromethyl)pyridin-3-yl)-2-iodoacetamide(Intermediate ED) (638 mg, 1.41 mmol) were mixed in 1,4-dioxane (5 mL)and DIPEA (616 μL, 3.52 mmol) was added. The mixture was stirred at 80°C. for 1.5 hours. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL)were added. The aqueous layer was extracted twice with DCM (2×10 mL).The combined organic layers were dried through a phase separator andconcentrated under reduced pressure. The crude product was dissolved inDCM and MeOH and SiliaMetS®Thiol was added. The mixture was stirred at40° C. for 1 hour and then filtered. The cake was washed with DCM. Thefiltrate was concentrated under reduced pressure to give the titlecompound.

LC-MS: Rt=1.17 min; MS m/z [M+H−Boc]⁺ 561.3, m/z [M−H]⁻ 659.3; UPLC-MS 3

Step 2: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((5-fluoro-2-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-4-(2-((5-fluoro-2-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(1.60 g, 1.45 mmol),2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(468 mg, 2.23 mmol) and XPhos Pd G3 (61.0 mg, 73.0 μmol) were mixed in1,4-dioxane (5 mL) and K₃PO₄ 1M in water (4.35 mL, 4.35 mmol) was added.The mixture was stirred at 80° C. for 2.5 hours. Water (10 mL), aq satNaHCO₃ (10 mL) and DCM (10 mL) were added. The aqueous layer wasextracted with DCM (2×10 mL). The combined organic layers were driedthrough a phase separator and concentrated under reduced pressure togive the title compound.

LC-MS: Rt=1.15 min; MS m/z [M+H−Boc]⁺ 565.4, m/z [M−H]⁻ 663.4; UPLC-MS 3

Step 3:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((5-fluoro-2-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(1.31 g, 1.58 mmol) was dissolved in DCM (5 mL) and TFA (1.00 mL, 13.0mmol) was added. The mixture was stirred at 40° C. for 20 minutes andthen concentrated under reduced pressure. Water (10 mL), aq sat NaHCO₃(10 mL) and EtOAc (10 mL) were added. The aqueous layer was extractedwith EtOAc (2×10 mL). The combined organic layers were dried through aphase separator and concentrated under reduced pressure to give thetitle compound.

LC-MS: Rt=0.84 min; MS m/z [M+H]⁺ 565.3, m/z [M−H]⁻ 563.1; UPLC-MS 3

Intermediate AS:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-bromo-5-ethyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (4.11 g, 8.37 mmol) and2-bromo-N-(4-(trifluoromethyl)phenyl)acetamide (2.36 g, 8.37 mmol) weredissolved in 1,4-dioxane (40 mL) and DIPEA (3.65 mL, 20.9 mmol) wasadded. The mixture was stirred at 80° C. for 4.5 hours. Water, aq satNaHCO₃ and DCM were added, and the aqueous layer was extracted twicewith DCM. The combined organic layers were dried through a phaseseparator and concentrated under reduced pressure. The crude product waspurified by column chromatography (RediSep Column: Silica 120 g, eluentDCM:DCM/MeOH (9/1) 100:0 to 0:100). The product containing fractionswere combined and concentrated under reduced pressure. This material waspurified by reverse phase preparative HPLC (RP-HPLC acidic 1: 20 to 80%B in 25 min with a plateau at 80% for 1 min). The product containingfractions were combined, basified with aq sat NaHCO₃, extracted twicewith DCM, dried through a phase separator and concentrated under reducedpressure to give the title compound.

LC-MS: Rt=1.22 min; MS m/z [M+H−Boc]⁺ 528.1, m/z [M−H]⁻ 626.0; UPLC-MS 4

Step 2: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(255 mg, 406 μmol),2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(128 mg, 609 μmol) and XPhos Pd G3 (3.43 mg, 4.06 μmol) were mixed in1,4-dioxane (4 mL). Then K₃PO₄ 1M in water (2.03 mL, 2.03 mmol) wasadded. The mixture was stirred at 80° C. for 4.5 hours. Water, aq satNaHCO₃ and DCM were added, and the aqueous layer was extracted twicewith DCM. The combined organic layers were dried through a phaseseparator. SiliaMetS®Thiol was added and the mixture was stirred at 40°C. for 2 hours. This mixture was filtered, and the solids were washedwith DCM. The filtrate was concentrated under reduced pressure, and thecrude product was purified by column chromatography (RediSep Column:Silica 12 g, eluent DCM:DCM/MeOH (9/1) 100:0 to 0:100). The productcontaining fractions were combined, concentrated and dried under HV togive the title compound.

LC-MS: Rt=1.14 min; MS m/z [M+H]⁺ 632.3, m/z [M−H]⁻ 630.1; UPLC-MS 4

Step 3:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(290 mg, 436 μmol) was dissolved in DCM (2 mL) and TFA (168 μL, 2.18mmol) was added. The mixture was stirred at RT for 1 hour. Water, aq satNaHCO₃ and DCM were added, and the aqueous layer was extracted with DCMtwice. The combined organic layers were dried through a phase separatorand concentrated under reduced pressure to give the title compound.

LC-MS: Rt=0.79 min; MS m/z [M+H]⁺ 532.2, m/z [M−H]⁻ 530.0; UPLC-MS 4

Intermediate AT:2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-bromo-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To the stirred solution of tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (2.50 g, 5.85 mmol) in DMF (20 mL) were added2-bromo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (IntermediateDP) (2.08 g, 7.02 mmol) and DIPEA (2.55 mL, 14.6 mmol) at 0° C. The RMwas stirred at RT for 16 hours. The RM was concentrated and ice coldwater was added. The precipitate was filtered off and dried under HV.The crude product was purified by column chromatography (Silica gelcolumn: Silica 40 g, eluent DCM:MeOH 100:0 to 90:10). The productcontaining fractions were combined, concentrated and dried under HV togive the title compound.

LC-MS: Rt=1.64 min; MS m/z [M+H]⁺ 642.2; UPLC-MS 11

Step 2: tert-butyl4-(5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-2-(pyridin-3-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To the stirred solution of tert-butyl4-(2-bromo-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(400 mg, 623 μmol) in 1,4-dioxane (3 mL) were added2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(153 mg, 1.25 mmol) and K₃PO₄ (1.87 mL, 1.87 mmol) at RT, and themixture was degassed for 2 minutes. XPhos Pd G3 (26.3 mg, 31.0 μmol) wasadded and the RM was degassed for 5 minutes, then it was stirred at 110°C. in the MW for 2 hours. The RM was concentrated and ice cold water wasadded. The solid was filtered and dried under HV. The solid wasdissolved in a small amount of 10% MeOH in DCM, filtered and dried.Smopex®-301 (350 mg) and DCM (50 mL) were added and the mixture wasstirred at RT for 16 hours. The suspension was filtered and the filtratewas concentrated under reduced pressure to give the title compound.

LC-MS: Rt=1.58 min; MS m/z [M+H]⁺ 641.3; UPLC-MS 11

Step 3:2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To the stirred solution of tert-butyl4-(5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-2-(pyridin-3-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(200 mg, 231 μmol) in DCM (10 mL) was added HCl 4M in 1,4-dioxane (4.00mL, 16.0 mmol) at 0° C. The RM was stirred at RT for 6 hours. The RM wasconcentrated under reduced pressure to give the title compound.

LC-MS: Rt=1.31 min; MS m/z [M+H]⁺ 541.3; UPLC-MS 11

Intermediate AU:2-(5-ethyl-2-(6-methylpyridin-3-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-bromo-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To the stirred solution of tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (16.9 g, 37.6 mmol) and2-iodo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (Intermediate DV)(15.5 g, 45.1 mmol) in 1,4-dioxane (200 mL) was added DIPEA (19.7 mL,113 mmol) at RT and the RM was stirred at 80° C. for 4 hours. The RM wasconcentrated, diluted with EtOAc and water, and extracted once withEtOAc. The organic layer was washed with brine, dried over Na₂SO₄ andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (RediSep Column: Silica 120 g, eluentheptane:EtOAc 70:30 to 5:95). The product containing fractions werecombined, concentrated, triturated in Et₂O and dried under HV to givethe title compound.

LC-MS: Rt=1.33 min; MS m/z [M+H−Boc]⁺ 542.2, m/z [M−H]⁻ 640.3; UPLC-MS 6

Step 2: tert-butyl4-(5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(6-methylpyridin-3-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(200 mg, 296 μmol), (6-methylpyridin-3-yl)boronic acid (51.2 mg, 355μmol), Na₂CO₃ (62.7 mg, 591 μmol),mesylate[(di(1-adamantyl)-n-butylphosphine)-2-(2-amino-1,1-biphenyl)]palladium(II)(22.7 mg, 30.0 μmol), 1,4-dioxane (657 μL) and water (329 μL) weremixed. The RM was degassed and purged with argon several times and thenstirred at 120° C. for 1 hour under MW irradiation (normal absorption).(6-Methylpyridin-3-yl)boronic acid (51.2 mg, 355 μmol), Na₂CO₃ (62.7 mg,591 μmol) andmesylate[(di(1-adamantyl)-n-butylphosphine)-2-(2-amino-1,1-biphenyl)]palladium(II)(22.7 mg, 30.0 μmol) were added to the RM, which was then stirred at120° C. for 30 minutes under MW irradiation (normal absorption). The RMwas diluted with THF (4 mL) and 5N aq NH₄Cl (8 mL). The organic layerwas collected and the aq layer was extracted 4 times with EtOAc. Thecombined organic layers were dried over Na₂SO₄ and concentrated. Theresidue was dissolved in DCM. PL-BnSH MP-Resin (59.9 mg, 148 μmol) wasadded and the resulting suspension was stirred at RT for 1 hour, thenfiltered. The filtrate was concentrated and dried under vacuum. Thecrude product was purified by column chromatography (RediSep Column:Silica 12 g, eluent DCM:DCM/MeOH (9/1) 100:0 to 50:50). The productcontaining fractions were combined, concentrated and dried under HV togive the title compound.

LC-MS: Rt=1.29 min; MS m/z [M+H]⁺ 655.5, m/z [M−H]⁻ 653.5; UPLC-MS 1

Step 3:2-(5-ethyl-2-(6-methylpyridin-3-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To a suspension of tert-butyl4-(5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(6-methylpyridin-3-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(127 mg, 184 μmol) in DCM (500 μL) was added TFA (500 μL, 6.49 mmol).The solution was stirred at RT for 40 minutes. The RM was concentratedand dried under HV to give the title compound as a brown foam.

LC-MS: Rt=0.74 min; MS m/z [M+H]⁺ 555.3, m/z [M−H]⁻ 553.4; UPLC-MS 1

Intermediate AV:2-(2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-bromo-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To the stirred solution of tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (3.00 g, 7.02 mmol) in DMF (20 mL) were added2-bromo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (IntermediateDP) (3.12 g, 10.5 mmol) and DIPEA (3.68 mL, 21.1 mmol) at 0° C. and theRM was stirred at RT for 14 hours. The RM was diluted with water andextracted with 5% MeOH in DCM. The combined organic layers were washedwith aq sat NaHCO₃ and brine, dried over Na₂SO₄ and concentrated underreduced pressure to give the title compound.

LC-MS: Rt=1.64 min; MS m/z [M+H]⁺ 642.1; UPLC-MS 11

Step 2: tert-butyl4-(2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To a stirred solution of tert-butyl4-(2-bromo-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(400 mg, 623 μmol) in 1,4-dioxane (10 mL) and water (3 mL) were added2-(5,6-dihydro-1,4-dioxin-2-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(238 mg, 1.12 mmol) and K₃PO₄ (264 mg, 1.25 mmol). The RM was degassedwith nitrogen for 15 minutes, then XPhos Pd G2 (24.5 mg, 31.0 μmol) wasadded and the RM was stirred at 100° C. for 2 hours. Water (20 mL) wasadded and the mixture was extracted with EtOAc (2×20 mL). The combinedorganic layers were washed with brine (20 mL), dried over Na₂SO₄ andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (Silica gel column: Silica 12 g, eluent DCM:MeOH100:0 to 99:1). The product containing fractions were combined,concentrated and dried under HV to give the title compound.

LC-MS: Rt=1.84 min; MS m/z [M+H]⁺ 648.2; UPLC-MS 12

Step 3:2-(2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To the stirred solution of tert-butyl4-(2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(260 mg, 393 μmol) in Et₂O (10 mL) was added HCl 2M in Et₂O (10.0 mL,20.0 mmol) and the RM was stirred at RT for 6 hours. The RM wasconcentrated under reduced pressure and washed with Et₂O to give thetitle compound.

LC-MS: Rt=1.91 min; MS m/z [M+H]⁺ 548.5; UPLC-MS 13

Intermediate AW:2-(2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-bromo-5-ethyl-4-(2-((3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (555 mg, 1.30 mmol) and2-chloro-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate EE) (403 mg, 1.30 mmol) were mixed in 1,4-dioxane (5 mL)and DIPEA (454 μL, 2.60 mmol) was added. The mixture was stirred at 80°C. for 5 hours, overnight at RT and then again at 80° C. for 8 hours.Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL) were added. Theaqueous layer was extracted with DCM (2×10 mL), and the combined organiclayers were dried through a phase separator and concentrated underreduced pressure. The crude product was purified in 2 portions byreverse phase preparative HPLC (2×RP-HPLC acidic 1: 5 to 100% in 20min). The product containing fractions were combined, basified with aqsat NaHCO₃, extracted twice with DCM, dried through a phase separatorand concentrated under reduced pressure to give the title compound as abeige solid.

LC-MS: Rt=1.34 min; MS m/z [M+H−Boc]⁺ 560.2, m/z [M−H]⁻ 658.3; UPLC-MS 1

Step 2: tert-butyl4-(2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-4-(2-((3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-4-(2-((3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(650 mg, 846 μmol),2-(5,6-dihydro-1,4-dioxin-2-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(197 mg, 931 μmol) and XPhos Pd G3 (35.8 mg, 42.0 μmol) were mixed in1,4-dioxane (5 mL) and K₃PO₄ 1M in water (2.54 mL, 2.54 mmol) was added.The mixture was stirred at 80° C. for 1 hour. Water (10 mL), aq satNaHCO₃ (10 mL) and DCM (10 mL) were added. The aqueous layer wasextracted with DCM (2×10 mL), and the combined organic layers were driedthrough a phase separator and concentrated under reduced pressure. Thecrude product was dissolved in DCM and MeOH and SiliaMetS®Thiol wasadded. The mixture was stirred at 40° C. for 1 hour. Then it wasfiltered, and the cake was washed with DCM. The filtrate wasconcentrated under reduced pressure to give the title compound.

LC-MS: Rt=1.26 min; MS m/z [M+H−Boc]⁺ 566.3, m/z [M−H]⁻ 664.2; UPLC-MS 1

Step 3:2-(2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

Tert-butyl4-(2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-4-(2-((3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(831 mg, 986 μmol) was dissolved in DCM (5 mL) and TFA (500 μL, 6.49mmol) was added. The mixture was stirred at 40° C. for 3 hours,concentrated under reduced pressure, and the crude product was purifiedin 2 portions by reverse phase preparative HPLC (2×RP-HPLC acidic 1: 5to 100% in 20 min). The product containing fractions were combined,basified with aq sat NaHCO₃, extracted twice with DCM, dried through aphase separator and concentrated under reduced pressure to give thetitle compound.

LC-MS: Rt=0.79 min; MS m/z [M+H]⁺ 566.4, m/z [M−H]⁻ 564.2; UPLC-MS 1

Intermediate AX:2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-bromo-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

See Intermediate AT Step 1.

Step 2: tert-butyl4-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(550 mg, 856 μmol) was suspended in 1,4-dioxane (15 mL).Rac-2-(4-methoxycyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(Intermediate DH) (408 mg, 1.71 mmol), XPhos Pd G3 (36.2 mg, 43.0 μmol)and K₃PO₄ 1M in water (856 μL, 856 μmol) were added and the RM wasstirred at 80° C. for 48 hours. The RM was concentrated under reducedpressure, ice cold water was added and it was extracted with 5% MeOH inDCM, washed with brine, dried over Na₂SO₄ and concentrated under reducedpressure. The crude product was purified by column chromatography(Silica gel column: Silica 12 g, eluent DCM:MeOH 100:0 to 95:5). Theproduct containing fractions were combined, concentrated and dried underHV to give the title compound.

LC-MS: Rt=1.65 min; MS m/z [M+H]+ 674.3; UPLC-MS 11

Step 3:rac-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To the solution of rac-tert-butyl4-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(180 mg, 246 μmol) in 1,4-dioxane (10 mL) was added HCl 4M in1,4-dioxane (10.0 mL, 40.0 mmol) at 0° C. and the RM was stirred at RTfor 16 hours. The RM was concentrated under reduced pressure, washedwith Et₂O (2×10 mL) and dried under HV to give the title compound, asthe HCl salt.

LC-MS: Rt=1.36 min; MS m/z [M+H]+ 574.3; UPLC-MS 11

Intermediate AY:2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-bromo-5-ethyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (320 mg, 748 μmol) was dissolved in 1,4-dioxane (2 mL)and 2-bromo-N-(4-(trifluoromethyl)phenyl)acetamide (226 mg, 800 μmol)was added, followed by DIPEA (314 μL, 1.80 mmol). The mixture wasstirred at 80° C. for 5 hours. Water, aq sat NaHCO₃ and DCM were added.The aqueous layer was extracted twice with DCM. The combined organiclayers were dried through a phase separator and concentrated underreduced pressure to give the title compound.

LC-MS: Rt=1.24 min; MS m/z [M−H]⁻ 626.0; UPLC-MS 7

Step 2: rac-tert-butyl4-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(991 mg, 1.06 mmol) was dissolved in 1,4-dioxane (5 mL) andrac-2-(4-methoxycyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(Intermediate DH) (252 mg, 1.06 mmol) was added, followed by XPhos Pd G3(14.5 mg, 17.0 μmol) and K₃PO₄ 1M in water (2.00 mL, 2.00 mmol). Themixture was stirred at 80° C. for 3 hours. Water, aq sat NaHCO₃ and DCMwere added. The aqueous layer was extracted twice with DCM. The combinedorganic layers were dried through a phase separator and concentratedunder reduced pressure. The crude product was purified by columnchromatography (RediSep Column: Silica 40 g, eluent DCM:DCM/MeOH (9/1)100:0 to 0:100). The product containing fractions were combined,concentrated and dried under HV to give the title compound.

LC-MS: Rt=1.27 min; MS m/z [M+H]⁺ 660.3, m/z [M−H]⁻ 658.2; UPLC-MS 7

Step 3:rac-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

Rac-tert-butyl4-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(575 mg, 636 μmol) was dissolved in DCM (5 mL) and TFA (490 μL, 6.36mmol) was added. The mixture was stirred at RT for 1.5 hours and thenconcentrated under reduced pressure. Water, aq sat NaHCO₃ and DCM wereadded, and the aqueous layer was washed twice with DCM. The combinedorganic layers were dried through a phase separator and concentratedunder reduced pressure to give the title compound.

LC-MS: Rt=0.80 min; MS m/z [M+H]⁺ 560.3, m/z [M−H]⁻ 558.1; UPLC-MS 4

Intermediate AZ: ethyl2-(4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)piperazin-1-yl)acetateStep 1: tert-butyl4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (500 mg, 1.17 mmol) was dissolved in DMF (1 mL) andDIPEA (613 μL, 3.51 mmol) was added at 0° C.2-Bromo-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (IntermediateDT) (556 mg, 1.76 mmol) was added and the RM was stirred at RT for 16hours. The RM was diluted with water and extracted with EtOAc. The crudeproduct was purified by column chromatography (Silica gel column: Silica4 g, eluent hexane:EtOAc 100:0 to 90:10). The product containingfractions were combined and concentrated to give the title compound.

LC-MS: Rt=1.67 min; MS m/z [M+H]+ 662.1; UPLC-MS 11

Step 2: tert-butyl4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(4-(2-ethoxy-2-oxoethyl)piperazin-1-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(450 mg, 645 μmol) was dissolved in DMSO (2 mL) and DMF (2 mL). Ethyl2-(piperazin-1-yl)acetate (Intermediate DC) (333 mg, 1.94 mmol) and KOAc(380 mg, 3.87 mmol) were added and the RM was stirred at 120° C. for 16hours. The RM was diluted with water and the precipitate was filteredoff and washed with 10% EtOAc in hexane to give the title compound.

LC-MS: Rt=1.60 min; MS m/z [M+H]+ 754.5; UPLC-MS 12

Step 3: ethyl2-(4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)piperazin-1-yl)acetate

Tert-butyl4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(4-(2-ethoxy-2-oxoethyl)piperazin-1-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(400 mg, 530 μmol) was dissolved in 1,4-dioxane (5 mL) and HCl 4N in1,4-dioxane (20.0 mL, 80.0 mmol) was added and the RM was stirred at RTfor 2 hours. The RM was concentrated under reduced pressure and washedwith Et₂O to get the title compound.

LC-MS: Rt=1.28 min; m/z [M−H]⁻ 652.2 UPLC-MS 11

Intermediate BA:2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-bromo-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (300 mg, 702 μmol),2-iodo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (Intermediate DV)(301 mg, 878 μmol) and DIPEA (368 μL, 2.11 mmol) were dissolved in DMF(1.75 mL) and the RM was stirred at 50° C. for 6.5 hours.2-iodo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (Intermediate DV)(50.0 mg, 146 μmol) was added and the RM was stirred at 50° C. for 16hours. 2-lodo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate DV) (50.0 mg, 146 μmol) was added and the RM was stirredat 50° C. for 7.5 hours. Water was added and the mixture was extractedwith EtOAc (3×20 mL). The combined organic layers were washed with waterand brine, dried through a phase separator and concentrated underreduced pressure. The crude product was adsorbed onto Isolute andpurified by column chromatography (RediSep Column: Silica 40 g, eluentDCM:MeOH 100:0 to 95:05). The product containing fractions were combinedand concentrated to give the title compound as a pale brown solid.

LC-MS: Rt=1.19 min; MS m/z [M+H−Boc]⁺ 542.2, m/z [M−H]⁻ 640.3; UPLC-MS 4

Step 2: tert-butyl4-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(452 mg, 704 μmol),2-(3,4-dihydro-2H-pyran-6-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(222 mg, 1.06 mmol) and XPhos Pd G3 (30.4 mg, 35.0 μmol) were mixed anddegassed and purged with argon several times. 1,4-Dioxane (3.7 mL) andK₃PO₄ 1M in water (2.11 mL, 2.11 mmol) were added and the RM was stirredat 90° C. for 2 hours. The RM was concentrated under reduced pressure,and the residue was extracted with EtOAc (3×50 mL). The organic layerswere washed with water (2×10 mL) and brine (10 mL), dried through aphase separator and concentrated under reduced pressure. The residue wasdissolved in DCM and MeOH and ISOLUTE® Si-Thiol (2.50 g) was added. Themixture was stirred for 45 minutes, then it was filtered and washed withDCM and MeOH. The filtrate was concentrated, and the crude product wasadsorbed onto Isolute and purified by column chromatography (RediSepColumn: Silica 40 g, eluent DCM:MeOH 100:0 to 95:05). The productcontaining fractions were combined and concentrated to give the titlecompound as a pale brown solid.

LC-MS: Rt=1.18 min; MS m/z [M+H]⁺ 646.5, m/z [M−H]⁻ 644.4; UPLC-MS 4

Step 3:2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Tert-butyl4-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(326 mg, 505 μmol) was dissolved in DCM (3.4 mL) and TFA (778 μL, 10.1mmol) was added and the RM was stirred at RT for 1.3 hours. The RM wasconcentrated under reduced pressure. The residue was dissolved in DCMand toluene and was concentrated under reduced pressure (two times). Theresidue was dried under HV to give the title compound as a pale brownsolid.

LC-MS: Rt=0.92 min; MS m/z [M+H]⁺ 546.2, m/z [M−H]⁻ 544.1; UPLC-MS 3

Intermediate BB:2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-bromo-5-ethyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

See Intermediate AS Step 1.

Step 2: tert-butyl4-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(245 mg, 390 μmol),2-(3,4-dihydro-2H-pyran-6-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(123 mg, 585 μmol) and XPhos Pd G3 (3.30 mg, 3.90 μmol) were dissolvedin 1,4-dioxane (2 mL). Then K₃PO₄ 1M in water (1.95 mL, 1.95 mmol) wasadded. The mixture was stirred at 80° C. for 15.5 hours. Water, aq satNaHCO₃ and DCM were added. The aqueous layer was extracted twice withDCM. The combined organic layers were dried through a phase separatorand concentrated under reduced pressure. The crude product was purifiedby column chromatography (RediSep Column: Silica 12 g, eluentDCM:DCM/MeOH (9/1) 100:0 to 60:40). The product containing fractionswere combined and concentrated to give the title compound.

LC-MS: Rt=1.17 min; MS m/z [M+H]⁺ 632.3, m/z [M−H]⁻ 630.1; UPLC-MS 4

Step 3:2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

Tert-butyl4-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(185 mg, 284 μmol) was dissolved in DCM (2 mL) and TFA (500 μL, 6.49mmol) was added. The mixture was stirred at RT for 1 hour. Water, aq satNaHCO₃ and DCM were added. The aqueous layer was extracted twice withDCM. The combined organic layers were dried through a phase separatorand concentrated under reduced pressure. As the crude product stillcontained tert-butyl4-(2-bromo-5-ethyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate,the Suzuki reaction was restarted using conditions of step 2 (seeabove). Water, aq sat NaHCO₃ and DCM were added to the crude product,and the aqueous layer was extracted twice with DCM. The combined organiclayers were mixed with SiliaMetS®Thiol and stirred at 40° C. for 2hours. Then it was filtered, dried through a phase separator andconcentrated under reduced pressure to give the title compound.

LC-MS: Rt=0.83 min; MS m/z [M+H]⁺ 532.2, m/z [M−H]⁻ 530.3; UPLC-MS 4

Intermediate BC:(S)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-bromo-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To a stirred solution of tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (1.00 g, 2.34 mmol) in DMF (1 mL) were added2-bromo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (IntermediateDP) (970 mg, 3.28 mmol) and DIPEA (1.02 mL, 5.85 mmol) at 0° C. The RMwas stirred at RT for 16 hours. The RM was concentrated, ice cold waterwas added and the precipitate was filtered off and dried under HV. Thecrude product was purified by column chromatography (Silica gel column:Silica 24 g, eluent DCM:MeOH 100:0 to 90:10). The product containingfractions were combined, concentrated and dried under HV to get thetitle compound as a brown solid.

LC-MS: Rt=1.64 min; MS m/z [M+H]⁺ 642.1; UPLC-MS 11

Step 2: tert-butyl(S)-4-(5-ethyl-2-(3-fluoropiperidin-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To a stirred solution of tert-butyl4-(2-bromo-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(400 mg, 535 μmol) in DMSO (4 mL) were added (S)-3-fluoropiperidine.HCl(374 mg, 2.68 mmol) and KOAc (315 mg, 3.21 mmol) at 0° C. The RM wasstirred at 120° C. for 16 hours. (S)-3-fluoropiperidine.HCl (110 mg, 623μmol) was added and the RM was stirred at 120° C. for 16 hours. Ice coldwater was added and the resultant solid was collected and dried underHV. The crude product was purified by column chromatography (Silica gelcolumn: Silica 24 g, eluent DCM:MeOH 100:0 to 90:10). The productcontaining fractions were combined and concentrated under reducedpressure to give the title compound as a brown solid.

LC-MS: Rt=1.64 min; MS m/z [M+H]⁺ 665.3; UPLC-MS 11

Step 3:(S)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To the stirred solution of tert-butyl(S)-4-(5-ethyl-2-(3-fluoropiperidin-1-yl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(120 mg, 181 μmol) in DCM (10 mL) was added HCl 4M in 1,4-dioxane (1.00mL, 4.00 mmol) at 0° C. The RM was stirred at RT for 16 hours and thenconcentrated under reduced pressure. Et₂O (10 mL) was added, decantedand the precipitate was dried under HV to give the title compound, asthe HCl salt.

LC-MS: Rt=1.36 min; MS m/z [M+H]⁺ 565.3; UPLC-MS 11

Intermediate BD: tert-butyl(R)-4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylate

Tert-butyl(2R)-4-(1-ethoxy-1,3-dioxopentan-2-yl)-2-methylpiperazine-1-carboxylate(Intermediate EK) (16.8 g, 48.1 mmol) was dissolved in EtOH (45 mL) and3-bromo-1H-1,2,4-triazol-5-amine (Intermediate ER) (7.50 g, 43.7 mmol)was added, followed by H₃PO₄ (5.29 g, 45.9 mmol). The RM was stirred at90° C. for 44 hours. DIPEA (22.9 mL, 131 mmol) and Boc₂O (5.08 mL, 21.9mmol) were added. The RM was stirred at RT for 1.5 hours. The reactionwas quenched with water, diluted with EtOAc and concentrated underreduced pressure. The residue was extracted with EtOAc (3×500 mL), andthe organic layers were washed with water (2×50 mL) and brine (2×50 mL),dried through a phase separator and concentrated under reduced pressure.This material was suspended in Et₂O (1 L) and stirred at reflux for 20minutes. The suspension was filtered, and the mother liquor wasconcentrated under reduced pressure. This material was mixed with hexane(2×600 mL), stirred at 40° C., and then filtered. The cake was dissolvedin DCM and MeOH, concentrated again and adsorbed onto Isolute andpurified twice by column chromatography (RediSep Column: Silica 120 g,eluent DCM:MeOH 100:0 to 85:15) and (RediSep Column: Silica 80 g, eluentDCM:MeOH 100:0 to 85:15). The product containing fractions were combinedand concentrated under reduced pressure to give the title compound as abeige solid.

LC-MS: Rt=0.94 min; MS m/z [M+H−Boc]⁺ 341.1, m/z [M−H]⁻ 439.3; UPLC-MS 4

¹H NMR (400 MHz, DMSO-d₆) δ 13.27 (s, 1H), 4.19 (m, 1H), 3.76 (m, 1H),3.52 (m, 1H), 3.37 (m, 1H), 3.10 (m, 1H), 2.86 (m, 1H), 2.71 (m, 3H),1.44 (s, 9H), 1.26 (m, 3H), 1.21 (t, J=7.5 Hz, 3H)

Intermediate BE:(R)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl(R)-4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylate

To a solution of tert-butyl(R)-4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylate(Intermediate BD) (4.07 g, 9.21 mmol) andN-(2-chloro-4-(trifluoromethyl)phenyl)-2-iodoacetamide (Intermediate DL)(3.68 g, 10.1 mmol) in DMF (61 mL) was added DIPEA (4.83 mL, 27.6 mmol).The RM was stirred at 50° C. for 2.5 hours, then allowed to cool to 30°C., and water was added slowly. The aqueous mixture was extracted twicewith EtOAc. The combined organic layers were washed 3 times with brine,dried through a phase separator and concentrated under reduced pressure.The crude product was adsorbed onto Isolute and purified by columnchromatography (RediSep Column: Silica 120 g, eluent heptane:EtOAc 100:0to 0:100). The product containing fractions were combined, concentratedand dried under HV to give the title compound as an orange-brown solid.

LC-MS: Rt=1.40 min; MS m/z [M+H−Boc]⁺ 576.2, m/z [M−H]⁻ 674.4; UPLC-MS 1

Step 2: tert-butyl(R)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylate

To a stirred solution of tert-butyl(R)-4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylate(5.93 g, 7.71 mmol), K₃PO₄ (5.58 g, 26.3 mmol) and Pd(dppf)Cl₂.DCM (358mg, 438 μmol) in 1,4-dioxane (44 mL) and water (15 mL) was added at 80°C. a solution of2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(2.39 g, 11.4 mmol) in 1,4-dioxane (15 mL) dropwise over 5 minutes andthe RM was stirred at 80° C. for 1 hour. The RM was concentrated anddiluted with EtOAc and water. The aqueous layer was extracted twice withEtOAc, and the combined organic phases were washed three times withbrine, dried through a phase separator and concentrated under reducedpressure. The residue was suspended in EtOAc, sonicated until a finesuspension was formed and then filtered. The cake was washed with asmall amount of EtOAc. The filtrate was concentrated under reducedpressure. The residue was suspended in a small amount of EtOAc,sonicated until a fine suspension was formed and then filtered. The cakewas washed with a small amount of EtOAc. Both cakes were combined andtriturated in Et₂O, sonicated and then filtered. The cake was washedwith a small amount of Et₂O then dried under vacuum to give product. TheEtOAc and Et₂O filtrates were combined and concentrated to dryness. Theresidue was adsorbed onto Isolute and purified by column chromatography(RediSep Column: Silica 80 g, eluent heptane:EtOAc 100:0 to 0:100). Theproduct containing fractions were combined and concentrated to giveproduct. Both product samples were combined to give the title compoundas a light brown solid.

LC-MS: Rt=1.36 min; MS m/z [M+H−Boc]⁺ 580.3, m/z [M−H]⁻ 678.5; UPLC-MS 1

Step 3:(R)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

A solution of tert-butyl(R)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylate(4.08 g, 5.70 mmol) in HCl 4N in 1,4-dioxane (42.8 mL, 171 mmol) wasstirred at RT for 45 minutes. The RM was quenched slowly with aq satNaHCO₃, then 10% MeOH in DCM was added. The biphasic mixture was stirredat RT overnight. Both phases were separated, and the aqueous layer wasconcentrated and extracted twice with 10% MeOH in DCM. The combinedorganic layers were dried through a phase separator, concentrated anddried under HV to give the title compound as a beige solid.

LC-MS: Rt=0.88 min; MS m/z [M+H]⁺ 580.3, m/z [M−H]⁻ 578.4; UPLC-MS 1

Intermediate BF:(R)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl(R)-4-(2-bromo-5-ethyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylate

Tert-butyl(R)-4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylate(Intermediate BD) (2.50 g, 5.66 mmol) and2-bromo-N-(4-(trifluoromethyl)phenyl)acetamide (2.02 g, 6.80 mmol) weredissolved in DMF (15 mL). DIPEA (2.97 mL, 17.0 mmol) was added and theRM was stirred at 85° C. for 4.75 hours, then allowed to cool to RT. TheRM was slowly added dropwise to ice-water (40 mL). The productprecipitated, the water was decanted and the residue was extracted withEtOAc (3×80 mL). The organic layers were washed with water (2×20 mL) andbrine (2×20 mL), dried through a phase separator and concentrated underreduced pressure. The crude product was adsorbed onto Isolute andpurified by column chromatography (RediSep Column: Silica 120 g, eluentDCM:MeOH 100:0 to 85:15). The product containing fractions were combinedand concentrated under reduced pressure to give the title compound as abrown solid foam.

LC-MS: Rt=1.16 min; MS m/z [M+H−Boc]⁺ 542.1, m/z [M−H]⁻ 640.2; UPLC-MS 4

Step 2: tert-butyl(R)-4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylate

Tert-butyl(R)-4-(2-bromo-5-ethyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylate(3.82 g, 4.16 mmol),2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(1.31 g, 6.25 mmol) and XPhos Pd G3 (176 mg, 208 μmol) were purged withargon. 1,4-Dioxane (20 mL) and K₃PO₄ 1M in water (12.5 mL, 12.5 mmol)were added and the RM was stirred at 90° C. for 3.5 hours. The RM wasallowed to cool to RT and stirred at RT overnight. DMF was removed, andthe aqueous residue was extracted with EtOAc (3×50 mL). The organiclayers were washed with water (2×10 mL) and brine (2×5 mL), driedthrough a phase separator and concentrated under reduced pressure. Theresidue was dissolved in DCM and MeOH. ISOLUTE® Si-Thiol (3.20 g) wasadded and the suspension was stirred for 30 minutes. The suspension wasfiltered and washed with DCM and MeOH. The filtrate was concentratedunder reduced pressure. The crude product was adsorbed onto Isolute andpurified by column chromatography (RediSep Column: Silica 120 g, eluentDCM:MeOH 100:0 to 85:15). The product containing fractions were combinedand concentrated under reduced pressure. The solid was dissolved in Et₂Oand hexane was added. The Et₂O was removed under reduced pressure. Theprecipitate was filtered off, the cake was mixed with hot hexane andfiltered. The beige solid was adsorbed onto Isolute and purified bycolumn chromatography (RediSep Column: Silica 120 g, eluent DCM:MeOH100:0 to 90:10). The product containing fractions were combined andconcentrated under reduced pressure. The solid was dissolved in Et₂O andhexane was added. Et₂O was removed under reduced pressure. The resultingsuspension was filtered, the cake was washed with hexane and dried underHV to give the title compound.

LC-MS: Rt=1.20 min; MS m/z [M+H−Boc]⁺ 546.3, m/z [M−H]⁻ 644.4; UPLC-MS 4

Step 3:(R)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

Tert-butyl(R)-4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylate(150 mg, 232 μmol) was dissolved in DCM (1 mL) and TFA (358 μL, 4.65mmol) was added. The RM was stirred at RT for 2.3 hours. The RM wasconcentrated under reduced pressure. DCM and toluene were added andremoved under reduced pressure. The residue was dried under HV to givethe title compound as brown foam.

LC-MS: Rt=0.88 min; MS m/z [M+H]⁺ 546.2, m/z [M−H]⁻ 544.0; UPLC-MS 4

Intermediate BG: tert-butyl(S)-4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylate

Tert-butyl(3S)-4-(1-ethoxy-1,3-dioxopentan-2-yl)-3-methylpiperazine-1-carboxylate(Intermediate EL) (1.32 g, 3.85 mmol) was dissolved in EtOH (5 mL) and3-bromo-1H-1,2,4-triazol-5-amine (Intermediate ER) (600 mg, 3.50 mmol)was added, followed by H₃PO₄ (444 mg, 3.85 mmol). The RM was stirred at85° C. for 22.5 hours, then it was allowed to cool to RT. After 30minutes at RT the RM was quenched with water, diluted with EtOAc andconcentrated under reduced pressure. The residue was extracted withEtOAc (3×60 mL), and the organic layers were washed with water (3×10 mL)and brine (2×10 mL), dried through a phase separator and concentratedunder reduced pressure. The crude product was adsorbed onto Isolute andpurified by column chromatography (RediSep Column: Silica 80 g, eluentDCM:DCM/MeOH (8/2) 100:0 to 0:100. The product containing fractions werecombined and concentrated under reduced pressure to give the titlecompound as an orange oil.

LC-MS: Rt=0.98 min; MS m/z [M+H]⁺ 441.3, m/z [M−H]⁻ 439.1; UPLC-MS 1

Intermediate BH:(S)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(2-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl(S)-4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylate

Tert-butyl(S)-4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylate(Intermediate BG) (236 mg, 417 μmol) andN-(2-chloro-4-(trifluoromethyl)phenyl)-2-iodoacetamide (Intermediate DL)(185 mg, 459 μmol) were dissolved in DMF (2.5 mL). DIPEA (219 μL, 1.25mmol) was added and the RM was stirred at 65° C. for 2.5 hours. The RMwas cooled to RT and water (5 mL) was added dropwise. The RM wasextracted with EtOAc (3×40 mL), and the organic layers were washed withwater (2×10 mL) and brine (2×10 mL), dried through a phase separator andconcentrated under reduced pressure. The crude product was dissolved inEt₂O and heptane was added. The Et₂O was evaporated and the suspensionwas filtered and washed with heptane. The cake was mixed with Et₂O againand heptane was added. Et₂O was removed and the suspension was filteredto give the title compound as a brown solid.

LC-MS: Rt=1.38 min; MS m/z [M+H−Boc]⁺ 576.1, m/z [M−H]⁻ 674.2; UPLC-MS 1

Step 2: tert-butyl(S)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylate

Tert-butyl(S)-4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylate(130 mg, 192 μmol), K₃PO₄ (122 mg, 576 μmol) and Pd(dppf)Cl₂.DCM (7.84mg, 9.60 μmol) were mixed under argon, then degassed and purged withargon several times. 1,4-Dioxane (3.2 mL) and water (1.6 mL) were added,followed by2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(60.5 mg, 288 μmol). The RM was stirred at 80° C. for 1 hour.Pd(dppf)Cl₂.DCM was added and the RM was stirred at 80° C. for 2 hours.Pd(dppf)Cl₂.DCM (5.00 mg, 6.12 μmol), K₃PO₄ (30.0 mg, 142 μmol) and2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(20.0 mg, 95.2 μmol) were added and the RM was stirred at 80° C. for 30minutes. The RM was cooled to RT and extracted with EtOAc (3×40 mL). Theorganic layers were washed with water (2×15 mL) and brine (15 mL), driedthrough a phase separator and concentrated under reduced pressure. Thecrude product was adsorbed onto Isolute and purified by columnchromatography (RediSep Column: Silica 40 g, eluent heptane:EtOAc 100:0to 20:80). The product containing fractions were combined andconcentrated to give the title compound as a white solid.

LC-MS: Rt=1.34 min; MS m/z [M+H−Boc]⁺ 580.3, m/z [M−H]⁻ 678.3; UPLC-MS 1

Step 3:(S)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(2-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl(S)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylate(53.7 mg, 79.0 μmol) was dissolved in DCM (600 μL) and TFA (122 μL, 1.58mmol) was added. The RM was stirred at RT for 1 hour and thenconcentrated under reduced pressure. DCM was added and removed twice.The residue was dried under HV to give the title compound as a brownsolid.

LC-MS: Rt=0.76 min; MS m/z [M+H]⁺ 580.3, m/z [M−H]⁻ 578.2; UPLC-MS 1

Intermediate BI: tert-butyl(R)-4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylate

Tert-butyl(3R)-4-(1-ethoxy-1,3-dioxopentan-2-yl)-3-methylpiperazine-1-carboxylate(Intermediate EM) (1.60 g, 4.66 mmol) was dissolved in EtOH (7 mL).3-bromo-1H-1,2,4-triazol-5-amine (Intermediate ER) (800 mg, 4.66 mmol)and H₃PO₄ (564 mg, 4.90 mmol) were added and the RM was stirred at 85°C. for 23.5 hours. The RM was cooled to RT. DIPEA (2.44 mL, 14.0 mmol)and Boc₂O (509 mg, 2.33 mmol) were added and the RM was stirred at RTfor 2 hours. The RM was quenched with a small amount of water, EtOH wasremoved under reduced pressure, and the residue was extracted with EtOAc(3×60 mL). The organic layers were washed with water (2×30 mL) and brine(2×25 mL), dried through a phase separator and concentrated underreduced pressure. The crude product was adsorbed onto Isolute andpurified by column chromatography (RediSep Column: Silica 120 g, eluentDCM:DCM/MeOH (8/2) 100:0 to 70:30). The product containing fractionswere combined and concentrated under reduced pressure. This material wasadsorbed onto Isolute and purified by column chromatography (RediSepColumn: Silica 80 g, eluent heptane:EtOAc 100:0 to 25:75). The productcontaining fractions were combined and concentrated under reducedpressure to give the title compound as a pale yellow solid.

LC-MS: Rt=0.97 min; MS m/z [M+H]⁺ 441.2, m/z [M−H]⁻ 439.2; UPLC-MS 1

Intermediate BJ:(R)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(2-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl(R)-4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylate

Tert-butyl(R)-4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylate(Intermediate BI) (191 mg, 433 μmol) andN-(2-chloro-4-(trifluoromethyl)phenyl)-2-iodoacetamide (Intermediate DL)(182 mg, 476 μmol) were dissolved in 1,4-dioxane (2.5 mL). DIPEA (227μL, 1.30 mmol) was added and the RM was stirred at 80° C. for 3.5 hours.The RM was cooled to RT, quenched with water (5 mL) and extracted withEtOAc (3×40 mL). The organic layers were washed with water (2×10 mL) andbrine (2×10 mL), dried through a phase separator and concentrated underreduced pressure. The crude product was adsorbed onto Isolute andpurified by column chromatography (RediSep Column: Silica 40 g, eluentheptane:EtOAc 100:0 to 45:55). The product containing fractions werecombined and concentrated under reduced pressure to give the titlecompound as a pale yellow solid.

LC-MS: Rt=1.38 min; MS m/z [M+H−Boc]⁺ 576.1, m/z [M−H]⁻ 674.2; UPLC-MS 1

Step 2: tert-butyl(R)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylate

Tert-butyl(R)-4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylate(183 mg, 270 μmol), K₃PO₄ (115 mg, 541 μmol), Pd(dppf)Cl₂.DCM (22.1 mg,27.0 μmol) and2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(85.0 mg, 406 μmol) were mixed under argon. The mixture was degassed andpurged with argon several times. 1,4-Dioxane (4.5 mL) and water (2.25mL) were added and the RM was stirred at 80° C. for 6 hours. The RM wascooled to RT and extracted with EtOAc (3×40 mL). The organic layers werewashed with water (2×10 mL) and brine (2×10 mL) and dried through aphase separator. The aqueous layers were extracted with EtOAc (2×40 mL),and the organic layers were dried through a phase separator. Thecombined organic layers were concentrated under reduced pressure. Tothis mixture were added K₃PO₄ (172 mg, 812 μmol) and Pd(dppf)C₁₂.DCM(22.1 mg, 27.0 μmol) at RT under argon. The mixture was degassed andpurged with argon several times. 1,4-Dioxane (4.5 mL) and water (2.25mL) were added, followed by2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(85.0 mg, 406 μmol). The RM was stirred at 80° C. for 1.25 hours, thencooled to RT and extracted with EtOAc (3×40 mL). The organic layers werewashed with water (2×10 mL) and brine (2×10 mL), dried through a phaseseparator and concentrated under reduced pressure to one third ofvolume. ISOLUTE® Si-Thiol (300 mg) was added and the mixture was stirredfor 45 minutes. The suspension was filtered and concentrated underreduced pressure. The crude product was adsorbed onto Isolute andpurified by column chromatography (RediSep Column: Silica 24 g, eluentheptane:EtOAc 100:0 to 0:100). The product containing fractions werecombined and concentrated under reduced pressure to give the titlecompound as a colorless solid.

LC-MS: Rt=1.35 min; MS m/z [M+H−Boc]⁺ 580.6, m/z [M−H]⁻ 678.4; UPLC-MS 1

Step 3:(R)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(2-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl(R)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylate(108 mg, 159 μmol) was dissolved in DCM (1.1 mL) and TFA (245 μL, 3.18mmol) was added. The RM was stirred at RT for 1 hour, then it wasconcentrated under reduced pressure. The residue was dissolved in DCMand concentrated under reduced pressure again. This was performed twice,then it was dried under HV. The residue was extracted with DCM (3×40 mL)and the organic layers were washed with aq sat NaHCO₃ (15 mL), driedthrough a phase separator and concentrated under reduced pressure togive the title compound as a beige solid.

LC-MS: Rt=0.75 min; MS m/z [M+H]⁺ 580.3, m/z [M−H]⁻ 578.2; UPLC-MS 1

Intermediate BK: tert-butyl(R)-4-(2-bromo-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylate

A solution of 3-bromo-1H-1,2,4-triazol-5-amine (Intermediate ER) (1.49g, 9.14 mmol) and tert-butyl(2R)-4-(1-ethoxy-1,3-dioxobutan-2-yl)-2-methylpiperazine-1-carboxylate(Intermediate EO) (3.18 g, 8.71 mmol) in AcOH (5.23 mL, 91.0 mmol) wasstirred at 110° C. for 4 hours. The RM was allowed to cool to RT andstand overnight. Water was added and the resulting suspension wasstirred at RT for 30 minutes and then concentrated and dried undervacuum. The crude product was dissolved in DCM and washed with water,insoluble parts between the two phases were collected and dried undervacuum, and the aqueous layer was extracted twice with DCM. The combinedorganic layers were dried through a phase separator, concentrated underreduced pressure and dried. This crude product was purified by columnchromatography (RediSep Column: Silica 24 g, eluent DCM:DCM/MeOH (9/1)100:0 to 0:100). The product containing fractions were combined andconcentrated to give the title compound.

LC-MS: Rt=0.94 min; MS m/z [M+H]⁺ 427.1, m/z [M−H]⁻ 425.1; UPLC-MS 8 Theaqueous layer from above extraction contained debocylated product andwas concentrated and dried under vacuum to give an orange-brown solid.Together with the collected insoluble parts, this material (3.00 g, 7.79mmol) was suspended in EtOH (20 mL). DIPEA (4.32 mL, 24.7 mmol) andBoc₂O (1.91 mL, 8.24 mmol) were added. The RM was stirred at RT for 50minutes and concentrated, and the residue was dissolved in DCM andwashed with water. The aqueous layer was extracted twice with DCM. Thecombined organic layers were dried through a phase separator,concentrated under reduced pressure and dried. This material waspurified by column chromatography (RediSep Column: Silica 120 g, eluentDCM:DCM/MeOH (9/1) 100:0 to 0:100). The product containing fractionswere combined and concentrated under reduced pressure to give the titlecompound.

LC-MS: Rt=0.94 min; MS m/z [M+H]⁺ 427.2, m/z [M−H]⁻ 425.1; UPLC-MS 8

Intermediate BL:(R)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl(R)-4-(2-bromo-5-methyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylate

A solution of tert-butyl(R)-4-(2-bromo-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylate(Intermediate BK) (1.29 g, 2.71 mmol) in DMF (12 mL) was cooled to 0°C., and DIPEA (1.39 mL, 7.96 mmol) was added. After 1 hour stirring at0° C. 2-bromo-N-(4-(trifluoromethyl)phenyl)acetamide (914 mg, 2.92 mmol)was added. After 10 minutes, the ice bath was removed and the mixturewas stirred at RT for 20.5 hours. Water (60 mL) was added, and themixture was extracted once with DCM and once with DCM and a small amountof MeOH. The organic layers were combined, dried through a phaseseparator and concentrated under reduced pressure to give a dark brownoil. The crude product was purified by column chromatography (RediSepColumn: Silica 80 g, eluent heptane:EtOAc 100:0 to 0:100), and theproduct containing fractions were combined to give the title compound.

LC-MS: Rt=1.24 min; MS m/z [M+H]⁺ 628.1, m/z [M−H]⁻ 626.0; UPLC-MS 8

Step 2: tert-butyl(R)-4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylate

Tert-butyl(R)-4-(2-bromo-5-methyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylate(845 mg, 1.16 mmol), 3,6-dihydro-2H-pyran-4-boronic acid pinacol ester(304 mg, 1.45 mmol) and XPhos Pd G3 (48.9 mg, 58.0 μmol) were introducedinto a MW vial, which was then degassed and purged with argon severaltimes. DMF (7.7 mL) and K₃PO₄ 1M in water (2.31 mL, 2.31 mmol) wereadded. The resulting mixture was stirred at 60° C. for 7.25 hours. TheRM was diluted with DCM and washed twice with aq sat NaHCO₃. Thecombined aqueous layer was extracted four times with DCM. The organiclayers were combined, dried through a phase separator and concentratedunder reduced pressure, and the crude product was purified by columnchromatography (RediSep Column: Silica 40 g, eluent heptane:EtOAc 100:0to 0:100). The product containing fractions were combined, concentratedand dried under HV. This material was dissolved in DCM and PL-BnSHMP-Resin (101 mg, 250 μmol) was added. The suspension was stirred at 37°C. for 1 hour and then filtered. The filtrate was concentrated and driedunder vacuum to give the title compound as a beige foam.

LC-MS: Rt=1.22 min; MS m/z [M+H]⁺ 632.3, m/z [M−H]⁻ 630.1; UPLC-MS 8

Step 3:(R)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

To a solution of tert-butyl(R)-4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylate(693 mg, 1.04 mmol) in DCM (1.6 mL) was added TFA (1.61 mL, 20.9 mmol).The RM was stirred at RT for 2 hours. The RM was concentrated todryness, and the crude product was dissolved in DCM/MeOH (1:1) and splitinto three portions. Those were filtered through PL-HCO₃ MP SPEcartridges (500 mg per 6 mL), prewashed with DCM/MeOH (1:1). Thecartridges were washed with DCM/MeOH (1:1), and the filtrates werecombined, concentrated and dried under vacuum. This material wasdissolved in DCM/MeOH (1:1), split into two portions and filteredthrough prewashed PL-HCO₃ MP SPE cartridges (500 mg per 6 mL). Thecartridges were washed with DCM/MeOH (1:1), and the filtrates werecombined, concentrated and dried under vacuum to give the title compoundas a light brown foam.

LC-MS: Rt=0.77 min; MS m/z [M+H]⁺ 532.3, m/z [M−H]⁻ 530.3; UPLC-MS 4

Intermediate BM: tert-butyl4-(2-bromo-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

3-Bromo-1H-1,2,4-triazol-5-amine (Intermediate ER) (4.00 g, 24.5 mmol),tert-butyl 4-(1-ethoxy-1,3-dioxobutan-2-yl)piperazine-1-carboxylate(Intermediate EP) (8.49 g, 27.0 mmol) and H₃PO₄ (2.97 g, 25.8 mmol) weremixed in EtOH (25 mL) and stirred at reflux for 18 hours. The RM wascooled to RT, DIPEA (12.9 mL, 73.6 mmol) and Boc₂O (1.71 mL, 7.36 mmol)were added, and the RM was stirred at RT for 1 hour. The RM was quenchedwith aq NH₄Cl, diluted with DCM, extracted twice with DCM, dried overNa₂SO₄, concentrated and dried. The crude product was crystallized fromDCM and TBME to give the title compound.

LC-MS: Rt=0.87 min; MS m/z [M+H]⁺ 413.1, m/z [M−H]⁻ 411.0; UPLC-MS 4

Intermediate BN:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-bromo-5-methyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate BM) (7.38 g, 17.9 mmol),2-bromo-N-(4-(trifluoromethyl)phenyl)acetamide (6.04 g, 21.4 mmol) andDIPEA (9.36 mL, 53.6 mmol) were dissolved in DMF (50 mL) and the RM wasstirred at 80° C. for 2 hours. The RM was cooled to RT, diluted withDCM, and the organic phase was extracted with aq sat NaHCO₃ and brine,dried over Na₂SO₄ and concentrated under reduced pressure. The crudeproduct was purified by column chromatography (eluent heptane:EtOAc/MeOH(9/1) 100:0 to 30:70). The product containing fractions were combinedand concentrated under reduced pressure and then crystallized from TBMEto give the title compound.

LC-MS: Rt=1.15 min; MS m/z [M+H]⁺ 614.0, m/z [M−H]⁻ 612.0; UPLC-MS 4

Step 2: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-methyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(7.70 g, 12.5 mmol),2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(3.95 g, 18.8 mmol), K₃PO₄ 1.5N (20.9 mL, 31.3 mmol) and XPhos Pd G3(1.06 g, 1.25 mmol) were dissolved in 1,4-dioxane (50 mL). The RM wasstirred at 90° C. for 1 hour and after cooling diluted with EtOAc. Theorganic phase was washed with aq sat NaHCO₃ and brine, dried over Na₂SO₄and concentrated under reduced pressure. This material was dissolved inDCM/MeOH (1:1) and ISOLUTE® Si-Thiol (258 mg) was added. After stirringfor 30 minutes, the mixture was filtered and concentrated. The crudeproduct was crystallized from DCM and TBME to give the title compound.

LC-MS: Rt=1.13 min; MS m/z [M+H]⁺ 618.2, m/z [M−H]⁻ 616.1; UPLC-MS 4

Step 3:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(5.95 g, 9.63 mmol) was dissolved in DCM (50 mL) and TFA (22.3 mL, 289mmol) was added. The RM was stirred at RT for 1 hour and thenconcentrated under reduced pressure. Toluene was added and removedagain, and this procedure was repeated. The residue was dissolved inEtOAc and washed with aq sat NaHCO₃ and brine. During the extraction theproduct crystallized, and the solids were collected and dried to givethe title compound.

LC-MS: Rt=0.84 min; MS m/z [M+H]⁺ 518.2, m/z [M−H]⁻ 516.0; UPLC-MS 4

Intermediate BO:2-(6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid Step 1: tert-butyl4-(2-bromo-4-(2-(tert-butoxy)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (23.0 g, 30.7 mmol) was dissolved in 1,4-dioxane (20mL) and tert-butyl 2-bromoacetate (6.79 mL, 46.0 mmol) was added,followed by DIPEA (16.1 mL, 92.0 mmol). The mixture was stirred at 80°C. for 3 hours. Water, aq sat NaHCO₃ and DCM were added. The aqueouslayer was extracted twice with DCM. The combined organic layers weredried through a phase separator and concentrated under reduced pressure.The crude product was suspended in Et₂O and sonicated for 5 minutes andfiltered. The cake was washed twice with Et₂O and dried under HV to givethe title compound. The filtrate was purified by column chromatography(RediSep Column: Silica 330 g, eluent DCM:DCM/MeOH (9/1) 100:0 to 80:20.The product containing fractions were combined, concentrated undervacuum and dried under HV and combined with the previously isolated cropto give the title compound.

LC-MS: Rt=1.13 min; MS m/z [M+H−Boc]⁺ 441.2/443.2, m/z [M−H]⁻539.2/541.2; UPLC-MS 3

Step 2: tert-butyl4-(4-(2-(tert-butoxy)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-4-(2-(tert-butoxy)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(9.12 g, 16.8 mmol),2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(4.86 g, 23.1 mmol) and XPhos Pd G3 (590 mg, 697 μmol) were mixed in1,4-dioxane (50 mL) and K₃PO₄ (50.5 mL, 50.5 mmol) was added. Themixture was stirred at 80° C. for 1 hour. Water, aq sat NaHCO₃ and DCMwere added. The aqueous layer was washed twice with DCM. The combinedorganic layers were dried through a phase separator and concentratedunder reduced pressure. The crude product was dissolved in DCM andSiliaMetS®Thiol was added. The mixture was stirred at RT for 1 hour thenfiltered. The cake was washed with DCM. The filtrate was concentratedunder reduced pressure to give the title compound.

LC-MS: Rt=1.10 min; MS m/z [M+H−Boc]⁺ 445.3, UPLC-MS 3

Step 3:2-(6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid

Tert-butyl4-(4-(2-(tert-butoxy)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(11.0 g, 17.8 mmol) was suspended in MeOH (50 mL) and NaOH 1M in water(26.7 mL, 26.7 mmol) was added. The suspension was stirred at RT for 1day. Water, aq 1M HCl and DCM were added. The aqueous layer wasextracted twice with DCM. The combined organic layers were dried througha phase separator and concentrated under reduced pressure. The crudeproduct was suspended in Et₂O, sonicated and filtered to give the titlecompound as a solid.

LC-MS: Rt=0.84 min; MS m/z [M+H−Boc]⁺ 389.3, m/z [M−H]⁻ 487.4; UPLC-MS 3

Intermediate BP:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-fluoro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

2-(6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid (Intermediate BO) (420 mg, 860 μmol) and2-fluoro-4-(trifluoromethyl)aniline (157 mg, 860 μmol) were mixed withEtOAc (6 mL). Et₃N (596 μL, 4.30 mmol) and T₃P 50% in EtOAc (1.02 mL,1.72 mmol) were added and the RM was stirred at RT for 1 hour. The RMwas extracted with EtOAc (3×40 mL), water (2×15 mL) and brine (15 mL).The organic layer was separated, filtered through a phase separator andconcentrated under reduced pressure. The crude product was adsorbed ontoIsolute and purified by column chromatography (RediSep Column: Silica 40g, eluent DCM:DCM/MeOH (8/2) 100:0 to 95:5). The product containingfractions were combined and concentrated to give the title compound as apale yellow solid.

LC-MS: Rt=1.18 min; MS m/z [M+H−Boc]⁺ 550.3, m/z [M−H]⁻ 648.4; UPLC-MS 4

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-fluoro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(346 mg, 533 μmol) was dissolved in DCM (3.6 mL) and TFA (821 μL, 10.7mmol) was added. The RM was stirred at RT for 1 hour, concentrated underreduced pressure. Residual TFA was removed under reduced pressure byazeotroping with DCM and toluene to give the title compound as a brightbrown foam as the trifluoroacetate salt.

LC-MS: Rt=0.86 min; MS m/z [M+H]⁺ 550.3, m/z [M−H]⁻ 548.3; UPLC-MS 4

Intermediate AJ:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

2-(6-(4-(Tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid (Intermediate BO) (150 mg, 307 μmol) and3-fluoro-2-methyl-4-(trifluoromethyl)aniline (65.0 mg, 337 μmol) wasdissolved in DCM (8 mL). Et₃N (170 μL, 1.23 mmol) was added followed byT₃P 50% in EtOAc (366 μL, 615 μmol). The yellow solution was stirred atRT for 5 days. T₃P 50% in EtOAc (2.00 mL, 3.36 mmol) and Et₃N (2.00 mL,14.4 mmol) were added and the mixture was stirred at RT for 15 minutes.Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL) were added. Theaqueous layer was extracted twice with DCM (2×10 mL). The combinedorganic layers was dried through a phase separator and concentratedunder reduced pressure to give the title compound.

LC-MS: Rt=1.29 min; MS m/z [M+H−Boc]⁺ 564.4, m/z [M−H]⁻ 662.3; UPLC-MS 1

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(335 mg, 242 μmol) was dissolved in DCM (10 mL) and TFA (1.00 mL, 13.0mmol) was added. The mixture was stirred at 40° C. for 30 minutes. Themixture was concentrated under reduced pressure. The crude product waspurified in 2 portions by reverse phase preparative HPLC (2×RP-HPLCacidic 1: 5 to 95% B in 20 min with a plateau at 95% for 1 min). Theproduct containing fractions were combined, basified with aq sat NaHCO₃,extracted twice with DCM, dried through a phase separator andconcentrated under reduced pressure to give the title compound.

LC-MS: Rt=0.83 min; MS m/z [M+H]⁺ 564.2, m/z [M−H]⁻ 562.3; UPLC-MS 1

Intermediate AM:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

2-(6-(4-(Tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid (Intermediate BO) (800 mg, 1.64 mmol) and5-fluoro-2-methyl-4-(trifluoromethyl)aniline (Intermediate DD) (396 mg,2.05 mmol) were mixed in DCM (5 mL). Then Et₃N (908 μL, 6.55 mmol) wasadded, followed by T₃P 50% in EtOAc (1.00 mL, 3.36 mmol). The mixturewas stirred at RT for 2 hours. T₃P 50% in EtOAc (1.00 mL, 3.36 mmol) wasadded and the mixture was stirred at RT for 1 hour. Water (10 mL), aqsat NaHCO₃ (10 mL) and DCM (10 mL) were added. The aqueous layer waswashed twice with DCM (2×10 mL). The combined organic layers were driedthrough a phase separator and concentrated under reduced pressure. Thecrude product was suspended in DCM and sonicated for 2 minutes andfiltered. The cake was washed with a small amount of DCM (2 mL) anddried under HV to give a solid as a first crop of the title compound.The filtrate was purified by column chromatography (RediSep Column:Silica 40 g, eluent DCM:DCM/MeOH (9/1) 100:0 to 50:50). The productcontaining fractions were combined, concentrated under vacuum, combinedwith the first crop of the title compound and dried under HV to affordthe title compound as a beige solid.

LC-MS: Rt=1.22 min; MS m/z [M+H]⁺ 664.4, m/z [M−H]⁻ 662.3; UPLC-MS 3

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(520 mg, 760 μmol) was dissolved in DCM (5 mL) and TFA (1.00 mL, 13.0mmol) was added. The mixture was stirred at 40° C. for 30 minutes. Themixture was concentrated under reduced pressure. The crude product waspurified in 2 portions by reverse phase preparative HPLC (2×RP-HPLCacidic 1: 20 to 50% B in 10 min with a plateau at 50% for 1 min). Theproduct containing fractions were combined, basified with aq sat NaHCO₃,extracted twice with DCM, dried through a phase separator andconcentrated under reduced pressure to give the title compound.

LC-MS: Rt=0.89 min; MS m/z [M+H]⁺ 564.3, m/z [M−H]⁻ 562.1; UPLC-MS 3

Intermediate AK:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

2-(6-(4-(Tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid (Intermediate BO) (4.20 g, 8.60 mmol) was dissolved in EtOAc (50mL) and Et₃N (4.77 mL, 34.4 mmol). 2-Chloro-4-(trifluoromethyl)aniline(1.68 g, 8.60 mmol) and T₃P 50% in DMF (10.2 mL, 17.2 mmol) were addedand the RM was stirred at RT for 1 hour. The RM was adsorbed ontoIsolute and purified by column chromatography (RediSep Column: Silica120 g, eluent cyclohexane:EtOAc 100:0 to 20:80). The product containingfractions were combined and concentrated to give the title compound.

LC-MS: Rt=1.35 min; MS m/z [M+H−Boc]⁺ 566.3/568.3, m/z [M−H]⁻664.4/666.4; UPLC-MS 1

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To a solution of tert-butyl4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(4.82 g, 7.24 mmol) in DCM (50 mL) was added TFA (8.36 mL, 109 mmol) andthe RM was stirred at RT for 2 hours. The RM was concentrated underreduced pressure. DCM was added and the mixture was extracted with aqNaHCO₃, adjusted to pH 10, extracted three times with DCM, dried througha phase separator and concentrated under reduced pressure to give thetitle compound.

LC-MS: Rt=0.79 min; MS m/z [M+H]⁺ 566.3/568.3, m/z [M−H]⁻ 564.4/566.4;UPLC-MS 1

Intermediate BQ:N-(2-chloro-5-fluoro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((2-chloro-5-fluoro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

2-(6-(4-(Tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid (Intermediate BO) (500 mg, 1.02 mmol) was dissolved in DCM (10 mL)and Et₃N (851 μL, 6.14 mmol).2-Chloro-5-fluoro-6-(trifluoromethyl)pyridin-3-amine (Intermediate DE)(263 mg, 1.13 mmol) and T₃P 50% in DMF (1.22 mL, 2.05 mmol) were addedand the RM was stirred at RT for 2 hours. The RM was adsorbed ontoIsolute and purified by column chromatography (RediSep Column: Silica 80g, eluent cyclohexane:EtOAc 100:0 to 0:100). The product containingfractions were combined and concentrated to give the title compound.

LC-MS: Rt=1.34 min; MS m/z [M+H]⁺ 685.3/687.3, m/z [M−H]⁻ 683.3/685.3;UPLC-MS 1

Step 2:N-(2-chloro-5-fluoro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((2-chloro-5-fluoro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(470 mg, 617 μmol) was dissolved in DCM (10 mL) and TFA (951 μL, 12.4mmol) was added. The RM was stirred at RT over the weekend. The RM wasadjusted to pH 10 by aq NaHCO₃, extracted twice with DCM, dried througha phase separator and concentrated under reduced pressure to give thetitle compound.

LC-MS: Rt=0.76 min; MS m/z [M+H]⁺ 585.2/587.2, m/z [M−H]⁻ 583.3/585.2;UPLC-MS 1

Intermediate BR:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(pentafluorosulfanyl)phenyl)acetamideStep 1: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-methyl-4-(pentafluorosulfanyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

2-(6-(4-(Tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid (Intermediate BO) (413 mg, 845 μmol) and2-methyl-4-(pentafluorosulfanyl)aniline (Intermediate DF) (249 mg, 930μmol) were mixed in DCM (10 mL). Et₃N (703 μL, 5.07 mmol) was added tothe suspension and it turned into an orange solution, then T₃P 50% inEtOAc (1.51 mL, 2.54 mmol) was added and the mixture was stirred at RTfor 1.5 hours. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL) wereadded. The aqueous layer was washed twice with DCM (2×10 mL). Thecombined organic layers were dried through a phase separator andconcentrated under reduced pressure to give the title compound.

LC-MS: Rt=1.32 min; MS m/z [M+H−Boc]⁺ 604.4, m/z [M−H]⁻ 702.5; UPLC-MS 1

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(pentafluorosulfanyl)phenyl)acetamide

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-methyl-4-(pentafluorosulfanyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(748 mg, 913 μmol) was dissolved in DCM (5 mL) and TFA (500 μL, 6.49mmol) was added. The mixture was stirred at 40° C. for 4 hours, then atRT overnight. The mixture was concentrated under reduced pressure. Thecrude product was purified by reverse phase preparative ISCO (RediSepColumn: C18 130 g, eluent water+0.1% TFA:ACN 100:0 to 0:100). Theproduct containing fractions were combined, basified with aq sat NaHCO₃,extracted twice with DCM, dried through a phase separator andconcentrated under reduced pressure to give the title compound.

LC-MS: Rt=0.89 min; MS m/z [M+H]⁺ 604.3, m/z [M−H]⁻ 602.2; UPLC-MS 1

Intermediate BS:N-(2,4-dichlorophenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((2,4-dichlorophenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To a solution of2-(6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid (Intermediate BO) (200 mg, 409 μmol) and 2,4-dichloroaniline (73.7mg, 450 μmol) in DMF (2 mL) were added Et₃N (170 μL, 1.23 mmol) and T₃P50% in DMF (487 μL, 819 μmol). The resulting brown solution was stirredat RT for 18 hours. The brown RM was diluted with DCM and extracted oncewith water. The aqueous layer was extracted twice with DCM and twicewith DCM/MeOH. The combined organic layers were dried through a phaseseparator, concentrated under reduced pressure and dried. The crudeproduct was purified by column chromatography (RediSep Column: Silica 12g, eluent DCM:DCM/MeOH (9/1) 100:0 to 60:40). The product containingfractions were combined and concentrated under reduced pressure to givethe title compound.

LC-MS: Rt=1.29 min; MS m/z [M+H−Boc]⁺ 532.2/534.2, m/z [M−H]⁻630.5/632.5; UPLC-MS 6

Step 2:N-(2,4-dichlorophenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To a solution of tert-butyl4-(4-(2-((2,4-dichlorophenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(220 mg, 313 μmol) in DCM (536 μL) was added TFA (536 μL, 6.96 mmol).The brown solution was stirred at RT for 70 minutes. The RM wasconcentrated under reduced pressure and dried under vacuum to give thetitle compound as the trifluoroacetate salt.

LC-MS: Rt=0.79 min; MS m/z [M+H]⁺ 532.3/534.3, m/z [M−H]⁻ 530.3/532.3;UPLC-MS 3

Intermediate BT:N-(5-chloro-2-methyl-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((5-chloro-2-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

2-(6-(4-(Tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid (Intermediate BO) (300 mg, 614 μmol) and5-chloro-2-methyl-6-(trifluoromethyl)pyridin-3-amine (Intermediate DG)(136 mg, 645 μmol) were mixed in EtOAc (4.2 mL). Et₃N (340 μL, 2.46mmol) and T₃P 50% in EtOAc (731 μL, 1.23 mmol) were added and the RM wasstirred at RT for 2.5 hours. The RM was quenched with water (5 mL) andit was extracted with EtOAc (3×50 mL), water (10 mL) and brine (10 mL).The organic layer was dried through a phase separator and concentratedunder reduced pressure. The crude product was adsorbed onto Isolute andpurified by column chromatography (RediSep Column: Silica 40 g, eluentDCM:MeOH 100:0 to 95:5). The product containing fractions were combinedand concentrated to give the title compound as a beige solid.

LC-MS: Rt=1.17 min; MS m/z [M+H−Boc]⁺ 581.4/583.4, m/z [M−H]⁻679.3/681.3; UPLC-MS 3

Step 2:N-(5-chloro-2-methyl-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((5-chloro-2-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(336 mg, 493 μmol) was dissolved in DCM (3.3 mL) and TFA (570 μL, 7.40mmol) was added. The RM was stirred at RT for 1.25 hours, then it wasconcentrated under reduced pressure. Traces of TFA were removed byazeotroping with DCM and toluene to give the title compoundtrifluoroacetate salt as a pale brown solid.

LC-MS: Rt=0.88 min; MS m/z [M+H]⁺ 581.4/583.4, m/z [M−H]⁻ 579.3/581.2;UPLC-MS 3

Intermediate BU:N-(4-chloro-2-methyl-5-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((4-chloro-2-methyl-5-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

2-(6-(4-(Tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid (Intermediate BO) (200 mg, 409 μmol) and4-chloro-2-methyl-5-(trifluoromethyl)aniline (90.0 mg, 430 μmol) weremixed in EtOAc (2.7 mL). Et₃N (227 μL, 1.64 mmol) and T₃P 50% in EtOAc(487 μL, 819 μmol) were added. The RM was stirred at RT for 1.25 hours,then it was quenched with water (5 mL). The RM was extracted with EtOAc(3×30 mL) then DCM (3×25 ml). The combined organic layers were washedsequentially with aq sat NaHCO₃ (5 mL), water (10 mL) and brine (2×5mL). The organic layer was dried through a phase separator andconcentrated under reduced pressure. The crude product was purified byreverse phase preparative HPLC (RP-HPLC acidic 1: 30 to 90% B in 20 minwith a plateau at 90% for 1 min). The product containing fractions werecombined and lyophilized to give the title compound as a white solid.

LC-MS: Rt=1.18 min; MS m/z [M+H−Boc]⁺ 580.3/582.3, m/z [M−H]⁻678.3/680.3; UPLC-MS 3

Step 2:N-(4-chloro-2-methyl-5-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((4-chloro-2-methyl-5-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(192 mg, 282 μmol) was dissolved in DCM (2 mL) and TFA (326 μL, 4.23mmol) was added. The RM was stirred at RT for 1.25 hours, then it wasconcentrated under reduced pressure. Traces of TFA were removed byazeotroping with DCM and toluene then it was dried under HV to give thetitle compound trifluoroacetate salt as a beige solid.

LC-MS: Rt=0.79 min; MS m/z [M+H]⁺ 580.3/582.2, m/z [M−H]⁻ 578.5/580.5;UPLC-MS 6 The free base of intermediate BU can be prepared bypartitioning the TFA salt between dichloromethane and saturated aqueousNaHCO₃, followed by separation and drying of the organic layer.

Intermediate BV:N-(5-chloro-4-ethyl-2-methylphenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((4-bromo-5-chloro-2-methylphenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

2-(6-(4-(Tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid (Intermediate BO) (1.00 g, 2.05 mmol) and4-bromo-5-chloro-2-methylaniline (515 mg, 2.34 mmol) were suspended inEtOAc (10 mL). Then Et₃N (1.14 mL, 8.79 mmol) was added, followed by T₃P50% in EtOAc (2.44 mL, 4.09 mmol). The suspension became a solution andit was stirred at RT for 1 hour. Water (10 mL), aq sat NaHCO₃ (10 mL)and EtOAc (10 mL) were added. The aqueous layer was extracted with EtOAc(2×10 mL). The combined organic layers were dried through a phaseseparator and concentrated under reduced pressure. The crude product wassuspended in DCM and Et₂O. The mixture was sonicated for 2 minutes andfiltered. The cake was washed with DCM and Et₂O. The cake was driedunder HV to give cake 1. The filtrate was concentrated under reducedpressure. The residue was suspended in MeOH and sonicated for 2 minutesand filtered. The cake was washed with Et₂O. The cake was dried under HVto give cake 2. The filtrate was purified by column chromatography(RediSep Column: Silica 40 g, eluent DCM:DCM/MeOH (9/1) 100:0 to 80:20).The product containing fractions were combined, concentrated undervacuum and dried under HV to afford a yellow solid. The yellow solid waspurified by reverse phase preparative HPLC (RP-HPLC acidic 1: 5 to 100%B in 20 min). The product containing fractions were combined, basifiedwith aq sat NaHCO₃, extracted twice with DCM, dried through a phaseseparator, concentrated under reduced pressure and combined with thecake 1 and cake 2 to give the title compound.

LC-MS: Rt=1.19 min; MS m/z [M+H−Boc]⁺ 590.3/592.3/594.3, m/z [M−H]⁻688.4/690.3/692.4; UPLC-MS 3

Step 2: tert-butyl4-(4-(2-((5-chloro-4-ethyl-2-methylphenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(4-(2-((4-bromo-5-chloro-2-methylphenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(908 mg, 1.25 mmol), potassium ethyltrifluoroborate (305 mg, 2.25 mmol),Pd(OAc)₂ (14.0 mg, 62.0 μmol), Cs₂CO₃ (1.22 g, 3.74 mmol) and RuPhos(58.2 mg, 125 μmol) were mixed in toluene (9.3 mL) and water (3.1 mL)and stirred at 100° C. for 6 hours. Then it was allowed to cool to RTovernight. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL) wereadded. The aqueous layer was extracted with DCM (2×10 mL). The combinedorganic layers were dried through a phase separator and concentratedunder reduced pressure. The crude product was dissolved in DCM and MeOHand SiliaMetS®Thiol was added. The mixture was stirred at 40° C. for 1hour and filtered. The cake was washed with DCM. The filtrate wasconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (RediSep Column: Silica 40 g, eluent DCM:DCM/MeOH(9/1) 100:0 to 80:20). The product containing fractions were combined,concentrated under vacuum and dried under HV to afford a pale beigesolid, which was further purified by SFC (SFC 1). The product containingfractions were combined and concentrated to give the title compound.

LC-MS: Rt=1.20 min; MS m/z [M+H−Boc]⁺ 540.4/542.4, m/z [M−H]⁻638.4/640.4; UPLC-MS 3

Step 3:N-(5-chloro-4-ethyl-2-methylphenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((5-chloro-4-ethyl-2-methylphenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(510 mg, 797 μmol) was dissolved in DCM (5 mL) and TFA (300 μL, 3.89mmol) was added. The mixture was stirred at 40° C. for 6 hours, left tostand at RT for 18 hours, then reheated to 40° C. and stirred for afurther 6 hours. The mixture was concentrated under reduced pressure.Water (20 mL), aq sat NaHCO₃ (20 mL) and DCM (20 mL) were added. Theaqueous layer was extracted with DCM (2×20 mL). The combined organiclayers were dried through a phase separator and concentrated underreduced pressure to give the title compound.

LC-MS: Rt=0.89 min; MS m/z [M+H]⁺ 540.4/542.4, m/z [M−H]⁻ 538.3/540.3;UPLC-MS 3

Intermediate BW:N-(4-chloro-2-methylphenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(2-bromo-4-(2-(tert-butoxy)-2-oxoethyl)-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate BM) (15.0 g, 36.3 mmol) was mixed with DMF (60.5 mL).Tert-butyl 2-bromoacetate (7.59 g, 38.1 mmol) and K₂CO₃ (10.0 g, 72.6mmol) were added. The RM was stirred at 65° C. for 3.3 hours, then itwas cooled to RT. Water (80 mL) was added and the suspension was stirredat RT for 30 minutes. The RM was cooled with an ice bath and continuedstirring for 2 hours. The suspension was filtered and washed with water.The cake was dried under HV to give the title compound as a beige solid.

LC-MS: Rt=1.13 min; MS m/z [M+H]⁺ 527.1/529.1, m/z [M−H]⁻ 525.0/527.0;UPLC-MS 4

Step 2: tert-butyl4-(4-(2-(tert-butoxy)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-4-(2-(tert-butoxy)-2-oxoethyl)-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(2.00 g, 3.79 mmol),2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(2.00 g, 9.52 mmol) and XPhos Pd G3 (160 mg, 190 μmol) were mixed in1,4-dioxane (10 mL) and K₃PO₄ 1M in water (9.48 mL, 9.48 mmol) wasadded. The mixture was stirred at 80° C. for 1 hour. Water, aq satNaHCO₃ and DCM were added. The aqueous layer were extracted twice withDCM. The combined organic layers were dried through a phase separatorand concentrated under reduced pressure. The crude product was dissolvedin DCM and SiliaMetS®Thiol was added. The mixture was stirred at 40° C.for 1 hour and filtered. The cake was washed with DCM. The filtrate wasconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (RediSep Column: Silica 80 g, eluent DCM:DCM/MeOH(9/1) 100:0 to 50:50). The product containing fractions were combinedand concentrated to give the title compound as a yellow oil.

LC-MS: Rt=1.10 min; MS m/z [M+H]⁺ 531.3; UPLC-MS 4

Step 3:2-(6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid

Tert-butyl4-(4-(2-(tert-butoxy)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(1.98 g, 3.62 mmol) was dissolved in THE (15 mL) and NaOH 1M in water(5.43 mL, 5.43 mmol) was added. The mixture was stirred at RT for 27hours. Water and DCM were added. The aqueous layer was washed twice withDCM. The combined organic layers were dried through a phase separatorand concentrated under reduced pressure. Then 1M HCl was added to theaqueous layer and it was extracted three times with DCM. The combinedorganic layers were dried through a phase separator and concentratedunder reduced pressure to give the title compound.

LC-MS: Rt=0.79 min; MS m/z [M+H−Boc]⁺ 375.3, m/z [M−H]⁻ 473.4; UPLC-MS 3

Step 4: tert-butyl4-(4-(2-((4-chloro-2-methylphenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

2-(6-(4-(Tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid (550 mg, 1.16 mmol) and 4-chloro-2-methylaniline (181 mg, 1.28mmol) were suspended in EtOAc (5 mL). Then T₃P 50% in EtOAc (2.76 mL,4.64 mmol) was added, followed by Et₃N (964 μL, 6.95 mmol). The yellowsolution was stirred at RT for 30 minutes. Water, aq sat NaHCO₃ andEtOAc were added. The aqueous layer were extracted twice with EtOAc. Thecombined organic layers was dried through a phase separator andconcentrated under reduced pressure to give the title compound.

LC-MS: Rt=1.11 min; MS m/z [M+H−Boc]⁺ 498.3/500.3, m/z [M−H]⁻596.4/598.3; UPLC-MS 4

Step 5:N-(4-chloro-2-methylphenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((4-chloro-2-methylphenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(1.02 g, 1.51 mmol) was suspended in DCM (5 mL) and TFA (1.00 mL, 13.0mmol) was added. The mixture was stirred at RT for 24 hours. Water, aqsat NaHCO₃ and DCM were added. The aqueous layer was extracted twicewith DCM. The combined organic layers were dried through a phaseseparator and concentrated under reduced pressure. The crude product wasdissolved in MeOH and DMF then stood at RT in an open flask for 3 hours.The resulting precipitate was filtered. The cake was washed with MeOHand dried under HV to give the title compound. The filtrate wasconcentrated under reduced pressure to give the title compound.

LC-MS: Rt=0.78 min; MS m/z [M+H]⁺ 498.3/500.3, m/z [M−H]⁻ 496.3/498.3;UPLC-MS 4

Intermediate BO:2-(6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid Step 1: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

See Intermediate AF.

Step 2: tert-butyl4-(4-(2-(tert-butoxy)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(3.00 g, 6.97 mol) and tert-butyl 2-bromoacetate (1.46 g, 7.32 mmol)were mixed in DMF (20 mL) under argon. DIPEA (3.65 mL, 20.9 mmol) wasadded and the RM was stirred at 55° C. for 4.5 hours. Water (50 mL) wasadded and the RM was stirred at RT overnight. The suspension wassonicated for 10 minutes, filtered and washed with water. The cake wasdried under HV, mixed with Et₂O and sonicated for 5 minutes. Thesuspension was stirred at reflux, filtered and the solid washed withEt₂O. A second crop of solid precipitates from the filtrate which wasfiltered. Both cakes were combined to give the title compound as a beigesolid.

LC-MS: Rt=1.16 min; MS m/z [M+H−Boc]⁺ 445.4, UPLC-MS 1

Step 3:2-(6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid

Tert-butyl4-(4-(2-(tert-butoxy)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(2.75 g, 5.00 mmol) was dissolved in THF (50 mL) and MeOH (20 mL). 1MNaOH (7.50 mL, 7.50 mmol) was added and the RM was stirred at RT for18.5 hours. The solvent was removed, water was added and the RM wasextracted with Et₂O (3×70 mL) and water (3×15 mL). The aqueous layer wascooled to 0° C. and 4M HCl (1.87 mL, 7.50 mmol) was added until pH 3.The resulting suspension was extracted with EtOAc (3×200 mL) and twicewith brine, then twice with EtOAc. The combined organic layers wereeluted through a phase separator and concentrated under reduced pressureto give the title compound as a bright brown solid.

LC-MS: Rt=0.76 min; MS m/z [M+H−Boc]⁺ 389.5, m/z [M−H]⁻ 487.2; UPLC-MS 1

Intermediate BX:N-(4-bromo-2-chlorophenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((4-bromo-2-chlorophenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

2-(6-(4-(Tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid (Intermediate BO) (250 mg, 512 μmol) and 4-bromo-2-chloroaniline(107 mg, 512 μmol) were mixed with EtOAc (3.6 mL). Et₃N (355 μL, 2.56mmol) and T₃P 50% in DMF (609 μL, 1.02 mmol) were added and the RM wasstirred at RT for 3 hours. The RM was extracted with EtOAc (3×40 mL),water (2×15 mL) and brine (2×10 mL). The combined organic layers wereeluted through a phase separator and concentrated under reducedpressure. The crude product was adsorbed onto Isolute and purified bycolumn chromatography (RediSep Column: Silica 40 g, eluent DCM:DCM/MeOH(8/2) 100:0 to 80:20). The product containing fractions were combinedand concentrated to give the title compound as a white solid.

LC-MS: Rt=1.27 min; MS m/z [M+H−Boc]⁺ 576.2/578.2/580.2, m/z [M−H]⁻674.3/676.3/678.3; UPLC-MS 1

Step 2:N-(4-bromo-2-chlorophenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((4-bromo-2-chlorophenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(265 mg, 391 μmol) was dissolved in DCM (2.6 mL) and TFA (603 μL, 7.83mmol) was added. The RM was stirred at RT for 1 hour, concentrated underreduced pressure. Residual TFA was removed by azeotoping with DCM underreduced pressure to give the title compound trifluoroacetate salt as awhite solid.

LC-MS: Rt=0.69 min; MS m/z [M+H]⁺ 576.1/578.1/580.2, m/z [M−H]⁻574.1/576.1/578.1; UPLC-MS 1

Intermediate BP:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-fluoro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

2-(6-(4-(Tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid (Intermediate BO) (250 mg, 512 μmol) and2-fluoro-4-(trifluoromethyl)aniline (94.0 mg, 512 μmol) were mixed withEtOAc (3.6 mL). Et₃N (355 μL, 2.56 mmol) and T₃P 50% in DMF (609 μL,1.02 mmol) were added and the RM was stirred at RT for 2.5 hours. The RMwas extracted with EtOAc (3×40 mL), water (2×15 mL) and brine (2×10 mL).The combined organic layers were eluted through a phase separator andconcentrated under reduced pressure. The crude product was adsorbed ontoIsolute and purified by column chromatography (RediSep Column: Silica 40g, eluent DCM:DCM/MeOH (8/2) 100:0 to 70:30). The product containingfractions were combined and concentrated to give the title compound as awhite solid.

LC-MS: Rt=1.28 min; MS m/z [M+H−Boc]⁺ 550.3, m/z [M−H]⁻ 648.3; UPLC-MS 1

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-fluoro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(237 mg, 365 μmol) was dissolved in DCM (2.5 mL) and TFA (562 μL, 7.30mmol) was added. The RM was stirred at RT for 1 hour, concentrated underreduced pressure. Residual TFA was removed by azeotroping with DCM twiceunder reduced pressure to give the title compound trifluoroacetate saltas a beige foam.

LC-MS: Rt=0.76 min; MS m/z [M+H]⁺ 550.3, m/z [M−H]⁻ 548.3; UPLC-MS 1

Intermediate BY:N-(2-bromo-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((2-bromo-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

2-(6-(4-(Tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid (Intermediate BO) (250 mg, 512 μmol) and2-bromo-4-(trifluoromethyl)aniline (123 mg, 512 μmol) were mixed inEtOAc (3.6 mL). Et₃N (355 μL, 2.56 mmol) and T₃P 50% in DMF (609 μL,1.02 mmol) were added and the RM was stirred at RT for 1.5 hours. The RMwas extracted with EtOAc (3×40 mL), water (2×15 mL) and brine (2×10 mL).The organic layers were eluted through a phase separator andconcentrated under reduced pressure. The crude product was placed ontoIsolute and purified by column chromatography (RediSep Column: Silica 40g, eluent DCM:DCM/MeOH (8/2) 100:0 to 70:30). The product containingfractions were combined and concentrated to give the title compound as awhite solid.

LC-MS: Rt=1.31 min; MS m/z [M+H−Boc]⁺ 610.2/612.2, m/z [M−H]⁻708.3/710.3; UPLC-MS 1

Step 2:N-(2-bromo-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((2-bromo-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(258 mg, 363 μmol) was dissolved in DCM (2.5 mL) and TFA (559 μL, 7.26mmol) was added. The RM was stirred at RT for 1 hour, then it wasconcentrated under reduced pressure. Residual TFA was removed byazeotroping with DCM under reduced pressure to give the title compoundtrifluoroacetate salt as a beige foam.

LC-MS: Rt=0.78 min; MS m/z [M+H]⁺ 610.3/612.3, m/z [M−H]⁻ 608.0/610.0;UPLC-MS 1

Intermediate BZ:2-(2-bromo-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-bromo-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To a stirred solution of tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (16.9 g, 37.6 mmol) and2-iodo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (Intermediate DV)(15.5 g, 45.1 mmol) in 1,4-dioxane (200 mL) was added DIPEA (19.7 mL,113 mmol) at RT and the RM was stirred at 80° C. for 4 hours. The RM wasconcentrated, diluted with EtOAc/water, extracted once with EtOAc andthe organic layer was washed with brine, dried over Na₂SO₄ andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (RediSep Column: Silica 120 g, eluentheptane:EtOAc 70:30 to 5:95). The product containing fractions werecombined, triturated with Et₂O and dried under HV to give the titlecompound as a white solid.

LC-MS: Rt=1.33 min; MS m/z [M+H−Boc]⁺ 542.2/544.2, m/z [M−H]⁻640.3/642.2; UPLC-MS 6

Step 2:2-(2-bromo-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To a stirred solution of tert-butyl4-(2-bromo-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(2.00 g, 3.11 mmol) in DCM (20 mL) was added TFA (10.0 mL, 130 mmol) atRT and the RM was stirred at RT for 5 minutes. The RM was diluted withDCM and NaHCO₃ and extracted 4 times with 10% MeOH in DCM. The organiclayer was dried over Na₂SO₄, concentrated and dried under HV to give thetitle compound as a beige foam.

LC-MS: Rt=0.74 min; MS m/z [M+H]⁺ 542.1/544.1, m/z [M−H]⁻ 540.4/542.4;UPLC-MS 6

Step 3:2-(2-bromo-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To a stirred solution of2-(2-bromo-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(1.00 g, 1.75 mmol) in DCM (20 mL) was added DIPEA (2.14 mL, 12.3 mmol),then 3-hydroxypicolinoyl chloride (Intermediate CV) (1.10 g, 7.01 mmol)in 4 portions at RT and the RM was stirred at RT for 1.5 hours. The RMwas diluted with DCM and water and extracted twice with DCM. Thecombined organic layers were washed with brine, dried over Na₂SO₄, andconcentrated. The crude product was purified by column chromatography(RediSep Column: Silica 40 g, eluent DCM:MeOH 100:0 to 50:50). Theproduct containing fractions were combined and concentrated to affordthe title compound as an off-white solid.

LC-MS: Rt=1.01 min; MS m/z [M+H]⁺ 663.2/665.2, m/z [M−H]⁻ 661.4/663.4;UPLC-MS 6

Intermediate CA:N-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

4H-1,2,4-triazol-3-amine (5.00 g, 59.5 mmol), tert-butyl4-(1-methoxy-1,3-dioxopentan-2-yl)piperazine-1-carboxylate (IntermediateEH) (20.6 g, 65.4 mmol) and H₃PO₄ (4.21 mL, 62.4 mmol) were mixed inEtOH (50 mL). The RM was stirred at reflux for 18 hours. H₃PO₄ (1.00 mL,14.8 mmol) was added and the RM was stirred at reflux for 18 hours. TheRM was concentrated under reduced pressure. AcOH (20 mL) was added andthe RM was stirred at 90° C. for 20 hours. The RM was concentrated underreduced pressure twice with toluene. THE was added, followed by DIPEA(31.2 mL, 178 mmol) and the RM was cooled to 0° C. Boc₂O (20.7 mL, 89.0mmol) was added and the RM was stirred for 7 hours. EtOAc was added andthe mixture was washed with citric acid and brine, dried over Na₂SO₄ andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (RediSep Column: Silica 120 g, eluent DCM:MeOH100:0 to 90:10). The product containing fractions were combined andconcentrated under reduced pressure. The product was crystallized fromDCM and TBME to give the title compound.

LC-MS: Rt=0.93 min; MS m/z [M+H]⁺ 349.2, m/z [M−H]⁻ 347.1; UPLC-MS 3

Step 2: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(IntAF)

The flask was flame-dried under reduced pressure and backfilled withnitrogen. The dry nitrogen flushed flask was charged with tert-butyl4-(5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(6.50 g, 18.5 mmol) and anhydrous THF (18.5 mL). To the white suspensionwas added zinc chloro 2,2,6,6-tetramethylpiperidide lithium chloridecomplex solution (45.1 mL, 59.1 mmol) with dropwise 1.5 mL/min. The RMwas stirred at RT for 2 hours. 4-bromo-3,6-dihydro-2H-pyran (6.02 mL,55.4 mmol), VPhos (958 mg, 1.85 mmol) and CPhos Pd G3 (1.57 g, 1.85mmol) were added. The RM was stirred at RT for 85.5 hours. The reactionwas quenched with aq. 5N NH₄Cl (100 mL). EtOAc (100 mL) was added andthe organic layer was collected. The aqueous layer was extracted withEtOAc (3×60 mL). The organic layers were combined, washed with 0.35% aq.NaHCO₃ (150 mL), brine (150 mL) and dried through a phase separator. Tothe filtrates was added ISOLUTE® Si-TMT (57.7 g, 27.7 mmol) and thesuspension was stirred at 40° C. for 1 hour. The mixture was filteredover a pad of celite. The filtrate was concentrated under reducedpressure. The crude product was adsorbed onto Isolute and purified bycolumn chromatography (Reveleris® HP silica cartridge 220 g, eluentDCM:MeOH 100:0 to 95:5). The product containing fractions were combinedand concentrated under reduced pressure. The residue was suspended inEt₂O and filtered. The cake was dried under HV to give the titlecompound as a white powder. The filtrate was concentrated under reducedpressure, adsorbed onto Isolute and purified again by columnchromatography (Reveleris® HP silica cartridge 120 g, eluent DCM:MeOH100:0 to 95:5). The product containing fractions were combined andconcentrated under reduced pressure to give a further crop of the titlecompound as a beige solid.

LC-MS: Rt=1.01 min; MS m/z [M+H]⁺ 431.3, m/z [M−H]⁻ 429.2; UPLC-MS 3

Step 3: tert-butyl4-(4-(2-((2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(200 mg, 465 μmol) and2-chloro-N-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)acetamide(Intermediate EF) (150 mg, 517 μmol) were mixed in 1,4-dioxane (1 mL)and DMF (1 mL). Then DIPEA (203 μL, 1.16 mmol) was added. The darksolution was stirred at 80° C. for 1 day. Water (10 mL), aq sat NaHCO₃(10 mL) and DCM (10 mL) were added. The aqueous layer was washed twicewith DCM (2×10 mL). The combined organic layers were dried through aphase separator and concentrated under reduced pressure. The crudeproduct was dissolved in MeOH and filtered. The filtrate was allowed tostand in an open flask for 2 days. The resulting solid was suspended inEt₂O (2 mL), sonicated, filtered and the cake was washed twice withEt₂O. The cake was dried under HV to give the title compound.

LC-MS: Rt=1.37 min; MS m/z [M+H−Boc]⁺ 584.3/586.3, m/z [M−H]⁻682.3/684.3; UPLC-MS 1

Step 4:N-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(165 mg, 229 μmol) was dissolved in DCM (2 mL) and TFA (100 μL, 1.30mmol) was added. The mixture was stirred at 40° C. for 2 hours, then atRT overnight and then again at 40° C. for 2 hours. The mixture wasconcentrated under reduced pressure and dried under HV to give the titlecompound trifluoroacetate salt.

LC-MS: Rt=0.85 min; MS m/z [M+H]⁺ 584.3/586.3, m/z [M−H]⁻ 582.3/584.4;UPLC-MS 1

Intermediate CB: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylateStep 1: tert-butyl4-(5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

4H-1,2,4-triazol-3-amine (13.6 g, 162 mmol) and tert-butyl4-(1-ethoxy-1,3-dioxobutan-2-yl)piperazine-1-carboxylate (IntermediateEP) (50.9 g, 162 mmol) were heated in AcOH (139 mL, 2.43 mol) at 100° C.for 70 minutes. The majority of the AcOH was removed in vacuo. Themixture was diluted with EtOH (100 mL) and heated at 88° C. for 18hours. The mixture was allowed to cool to RT and filtered. The solid waswashed with EtOH (120 mL) and dried in vacuo at 50° C. overnight to givethe title compound as an off-white solid.

LC-MS: Rt=0.79 min; MS m/z [M+H]⁺ 335.5, m/z [M−H]⁻ 333.4; UPLC-MS 8

Step 2: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

A flask was flame-dried under reduced pressure and backfilled withargon. Tert-butyl4-(5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(5.00 g, 14.7 mmol) was suspended in anhydrous THF (14.7 mL). To thewhite suspension was added at RT zinc chloro2,2,6,6-tetramethylpiperidide lithium chloride complex solution (35.9mL, 32.2 mmol) with dropwise 1 mL/min. The RM was stirred for 1.5 hours.Then it was stored in the freezer under argon overnight. The nextmorning it was stirred at RT for 4 hours. 4-bromo-3,6-dihydro-2H-pyran(3.18 mL, 29.3 mmol), CPhos (392 mg, 879 μmol) and CPhos Pd G3 (746 mg,879 μmol) were introduced under argon. The RM was stirred at RT for 68.5hours. The reaction was partitioned between THF (100 mL) and aq. NH₄Cl5N (100 mL). The aqueous layer was back-extracted with EtOAc (3×50 mL).The organic layers were combined, washed with aq. 10% Na₂S₂O₃ (150 mL),brine (150 mL) and dried through a phase separator. The solvent wascollected and treated with ISOLUTE® Si-TMT (36.6 g, 17.6 mmol). Thesuspension was stirred at 40° C. for 1 hour and was filtered off over apad of celite. Removal of volatiles under pressure. The residue wasadsorbed onto Isolute and purified by column chromatography (FlashPure®EcoFlex silica cartridge 330 g, eluent DCM:MeOH 100:0 to 95:5). Theproduct containing fractions were combined and concentrated to give thetitle compound as an off-white solid.

LC-MS: Rt=0.89 min; MS m/z [M+H]⁺ 417.3, m/z [M−H]⁻ 415.2; UPLC-MS 8

Intermediate BN:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate CB) (500 mg, 1.20 mmol)2-bromo-N-(4-(trifluoromethyl)phenyl)acetamide (406 mg, 1.44 mmol) andK₂CO₃ (232 mg, 1.68 mmol) were mixed in DMF (5 mL). The RM was stirredat 80° C. for 30 minutes. 2-Bromo-N-(4-(trifluoromethyl)phenyl)acetamide(40.6 mg, 144 μmol) was added again and the RM was stirred at 80° C. for30 minutes. The RM was diluted with EtOAc, washed with aq sat NaHCO₃ andbrine, dried over Na₂SO₄ and concentrated under reduced pressure. Thecrude product was crystallized from TBME, then dried under HV to givethe title compound.

LC-MS: Rt=1.19 min; MS m/z [M+H]⁺ 618.3, m/z [M−H]⁻ 616.2; UPLC-MS 8

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(600 mg, 971 μmol) was dissolved in DCM (5 mL). TFA (2.25 mL, 29.1 mmol)was added and the RM was stirred at RT for 1 hour. The RM wasconcentrated under reduced pressure, dissolved twice in toluene andconcentrated under reduced pressure again. The residue was dissolved inEtOAc, washed with aq sat NaHCO₃ and brine, dried over Na₂SO₄ andconcentrated under reduced pressure to give the title compound.

LC-MS: Rt=0.74 min; MS m/z [M+H]⁺ 518.2, m/z [M−H]⁻ 516.1; UPLC-MS 8

Intermediate CC:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl(2R,5S)-4-(3-amino-6-ethyl-2-imino-4-oxo-1,2,3,4-tetrahydropyrimidin-5-yl)-2,5-dimethylpiperazine-1-carboxylate

To a beige suspension of hydrazinecarboximidamide hydrochloride (2.95 g,26.7 mmol) in EtOH (44.4 mL) was added tetrabutylammonium hydroxide(18.3 mL, 28.0 mmol) at RT. The yellow turbid solution was stirred at50° C. for 2.75 hours. Tert-butyl(2R,5S)-4-(1-ethoxy-1,3-dioxopentan-2-yl)-2,5-dimethylpiperazine-1-carboxylate(Intermediate EN) (5.00 g, 13.3 mmol) was added and the RM was stirredat 80° C. for 17.5 hours. The EtOH was removed under vacuum. Water wasadded to the brown oily residue, which was then stirred for 1 hour.Extracted three times with EtOAc. The combined organic layers were driedover Na₂SO₄, concentrated and dried under vacuum to give a dark brownoily residue. The crude product was purified by column chromatography(Silica gel column: Silica 120 g, eluent DCM:DCM/MeOH (8/2) 100:0 to60:40). The product containing fractions were combined and concentratedto give the title compound as a beige foam.

LC-MS: Rt=0.88 min; MS m/z [M+H]⁺ 367.4, m/z [M−H]⁻ 365.4; UPLC-MS 1

Step 2: tert-butyl(2R,5S)-4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-dimethylpiperazine-1-carboxylate

To a solution of tert-butyl(2R,5S)-4-(3-amino-6-ethyl-2-imino-4-oxo-1,2,3,4-tetrahydropyrimidin-5-yl)-2,5-dimethylpiperazine-1-carboxylate(1.55 g, 3.98 mmol) in NMP (4.42 mL) was added3,6-dihydro-2H-pyran-4-carbaldehyde (Intermediate DK) (535 mg, 4.77mmol) then iron(III) chloride (645 mg, 3.98 mmol). The dark brownmixture was stirred at 50° C. for 18.5 hours.3,6-dihydro-2H-pyran-4-carbaldehyde (Intermediate DK) (150 mg, 1.34mmol) was added to the dark brown RM, which was then stirred at 50° C.for 5 hours. The RM was cooled down to RT. Water was added. The mixturewas stirred at RT overnight. No clean suspension obtained. It wasextracted three times with DCM. The combined organic layers were driedthrough a phase separator and concentrated under reduced pressure togive the title compound as a brown liquid.

LC-MS: Rt=1.01 min; MS m/z [M+H]⁺ 459.6, m/z [M−H]⁻ 457.4; UPLC-MS 1

Step 3: tert-butyl(2R,5S)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-dimethylpiperazine-1-carboxylate

To a dark brown solution of tert-butyl(2R,5S)-4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-dimethylpiperazine-1-carboxylate(6.06 g, 3.96 mmol) in 1,4-dioxane (26.4 mL) were addedN-(2-chloro-4-(trifluoromethyl)phenyl)-2-iodoacetamide (Intermediate DL)(1.60 g, 3.96 mmol), then DIPEA (900 μL, 5.15 mmol). The dark brownsolution was stirred at 50° C. for 5.25 hours. The dark brown mixturewas cooled down to RT. Water and EtOAc were added. The biphasic mixturewas vigorously stirred for 30 minutes. Both phases were separated. Theaqueous layer was extracted twice with EtOAc. The combined organiclayers were dried through a phase separator and concentrated underreduced pressure to give a dark brown residue. The crude product waspurified by column chromatography (Silica gel column: Silica 80 g,eluent heptane:EtOAc 100:0 to 0:100). The product containing fractionswere combined and concentrated under reduced pressure to give the titlecompound as a light brown solid.

LC-MS: Rt=1.38 min; MS m/z [M+H]⁺ 694.3/696.3, m/z [M−H]⁻ 692.3/694.3;UPLC-MS 1

Step 4:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

A light brown solution of tert-butyl(2R,5S)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-dimethylpiperazine-1-carboxylate(934 mg, 1.28 mmol) in 4N HCl in 1,4-dioxane (6.39 mL, 25.6 mmol) wasstirred at RT for 1 hour. The RM was dissolved in DCM and washed oncewith aq sat NaHCO₃. The aqueous layer was extracted once with DCM.Presence of beige solids at the bottom of the aqueous phase. The aqueouslayer was extracted twice with DCM/MeOH (95/5). The aqueous layer wasfiltered to give a beige solid and the filtrate extracted once more withDCM. The filtered beige solids were dissolved in DCM/MeOH. The turbidsolution was combined with the previous organic phases. They were driedthrough a phase separator, concentrated and dried under vacuum to givethe title compound hydrochloride salt.

LC-MS: Rt=0.82 min; MS m/z [M+H]⁺ 594.4/596.4, m/z [M−H]⁻ 592.4/594.4;UPLC-MS 1

Intermediate CD: tert-butyl4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,3-dimethylpiperazine-1-carboxylateStep 1: tert-butyl4-(3-amino-6-ethyl-2-imino-4-oxo-1,2,3,4-tetrahydropyrimidin-5-yl)-2,3-dimethylpiperazine-1-carboxylate

To a stirred solution of hydrazinecarboximidamide.HCl (13.6 g, 123 mmol)in EtOH (250 mL) was added tetrabutylammonium hydroxide (100 mL, 153mmol) at RT. The RM was stirred at 55° C. for 30 minutes. Tert-butyl4-(1-methoxy-1,3-dioxopentan-2-yl)-2,3-dimethylpiperazine-1-carboxylate(Intermediate EQ) (21.0 g, 61.3 mmol) was added and the RM was stirredat 90° C. for 16 hours. The RM was concentrated. The residue wasdissolved in DCM/water, extracted twice with DCM and the combinedorganic extracts were dried over Na₂SO₄ and concentrated. The crudeproduct was purified by column chromatography (Silica gel column: Silica220 g, eluent DCM:DCM/MeOH (8/2) 100:0 to 50:50). The product containingfractions were combined and concentrated under reduced pressure to givethe title compound as an oil.

LC-MS: Rt=0.87/0.91 min; MS m/z [M+H]⁺ 367.3; UPLC-MS 1

Step 2: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,3-dimethylpiperazine-1-carboxylate

To a stirred solution of tert-butyl4-(3-amino-6-ethyl-2-imino-4-oxo-1,2,3,4-tetrahydropyrimidin-5-yl)-2,3-dimethylpiperazine-1-carboxylate(5.00 g, 12.3 mmol) and 3,6-dihydro-2H-pyran-4-carbaldehyde(Intermediate DK) (1.62 g, 14.7 mmol) in NMP (10 mL) was added iron(III)chloride (3.98 g, 24.6 mmol) at RT and the RM was stirred at 55° C. for40 hours. The RM was diluted with DCM/water, filtered through celite,the filtrate was extracted twice with DCM and the combined organicextracts were dried over Na₂SO₄ and concentrated to give the titlecompound as a brown oil.

LC-MS: Rt=0.99 min; MS m/z [M+H]⁺ 459.4, m/z [M−H]⁻ 457.4; UPLC-MS 1

Step 3: tert-butyl4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,3-dimethylpiperazine-1-carboxylate

To a stirred solution of tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,3-dimethylpiperazine-1-carboxylate(9.00 g, 12.6 mmol) andN-(2-chloro-4-(trifluoromethyl)phenyl)-2-iodoacetamide (Intermediate DL)(5.02 g, 13.8 mmol) in 1,4-dioxane (100 mL) was added DIPEA (2.85 mL,16.3 mmol) at RT and the RM was stirred at 45° C. for 14 hours. The RMwas diluted with EtOAc/water, extracted twice with EtOAc and thecombined organic extracts were washed with water and brine, dried overNa₂SO₄ and concentrated. The crude product was purified by columnchromatography (Silica gel column: Silica 80 g, eluent heptane:EtOAc100:0 to 0:100). The product containing fractions were combined andconcentrated under reduced pressure to give the title compound as abeige solid.

LC-MS: Rt=1.36 min; MS m/z [M+H]⁺ 694.4/696.4, m/z [M−H]⁻ 692.5/694.5;UPLC-MS 1

Chiral separation of tert-butyl4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,3-dimethylpiperazine-1-carboxylate:

The compound was dissolved in 80 mL MeOH/DCM.

Preparative chiral SFC (instrument: MG II; column: ChiralPak IC, 250×30mm I.D., 5 μm; eluent: A=CO₂, B=MeOH+0.1% NH₄OH; flow rate: 50.0 mL/min;detection: 220 nm; column temperature: 38° C.; back pressure: 100 bar;injection volume: 2 mL; Gradient: 0 to 35% B

Intermediate CE: tert-butyl(2S,3R)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,3-dimethylpiperazine-1-carboxylateor tert-butyl(2R,3S)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,3-dimethylpiperazine-1-carboxylate

First Eluting Stereoisomer

Chiral HPLC (C-HPLC 9): Rt=1.649 min, 100% ee

LC-MS: Rt=1.38 min; MS m/z [M+H]⁺ 694.6/696.6, m/z [M−H]⁻ 692.4/694.4;UPLC-MS 1

Intermediate CF: tert-butyl(2S,3S)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,3-dimethylpiperazine-1-carboxylate

Second Eluting Stereoisomer

Chiral HPLC (C-HPLC 9): Rt=1.909 min, 99.8% ee

LC-MS: Rt=1.36 min; MS m/z [M+H]⁺ 694.5/696.5, m/z [M−H]⁻ 692.5/694.5;UPLC-MS 1

Intermediate CG: tert-butyl(2R,3S)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,3-dimethylpiperazine-1-carboxylateor tert-butyl(2S,3R)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,3-dimethylpiperazine-1-carboxylate

Third Eluting Stereoisomer

Chiral HPLC (C-HPLC 9): Rt=2.505 min, 99.8% ee

LC-MS: Rt=1.36 min; MS m/z [M+H]⁺ 694.5/696.5, m/z [M−H]⁻ 692.5/694.5;UPLC-MS 1

Intermediate CH: tert-butyl(2R,3R)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,3-dimethylpiperazine-1-carboxylate

Fourth Eluting Stereoisomer

Chiral HPLC (C-HPLC 9): Rt=3.202 min, 100% ee

LC-MS: Rt=1.38 min; MS m/z [M+H]⁺ 694.3/696.3, m/z [M−H]⁻ 692.3/694.3;UPLC-MS 1

Intermediate CI:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-((2R,3R)-2,3-dimethylpiperazin-1-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

HCl 4N in 1,4-dioxane (3.00 mL, 12.0 mmol) was added to tert-butyl(2R,3R)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,3-dimethylpiperazine-1-carboxylate(Intermediate CH) (717 mg, 950 μmol) at RT and the RM was stirred at RTfor 30 minutes. The RM was diluted with DCM/NaHCO₃, extracted twice withDCM and the combined organic extracts were washed with brine, dried overNa₂SO₄ and concentrated to give the title compound as a white solid.

LC-MS: Rt=0.85 min; MS m/z [M+H]⁺ 594.4/596.4, m/z [M−H]⁻ 592.4/594.5;UPLC-MS 1

Intermediate CJ:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-((2S,3S)-2,3-dimethylpiperazin-1-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

HCl 4N in 1,4-dioxane (21.3 μL, 85.0 μmol) was added to tert-butyl(2S,3S)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,3-dimethylpiperazine-1-carboxylate(Intermediate CF) (59.0 mg, 85.0 μmol) at RT and the RM was stirred atRT for 5 minutes. The RM was diluted with DCM/NaHCO₃, extracted twicewith DCM and the combined organic extracts were washed with brine, driedover Na₂SO₄ and concentrated to give the title compound as a whitesolid.

LC-MS: Rt=0.80 min; MS m/z [M+H]⁺ 594.6/596.6, m/z [M−H]⁻ 592.2/594.3;UPLC-MS 1

Intermediate CK:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-((2R,3S)-2,3-dimethylpiperazin-1-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideorN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-((2S,3R)-2,3-dimethylpiperazin-1-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

HCl 4N in 1,4-dioxane (1.00 mL, 4.00 mmol) was added to tert-butyl(2S,3R)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,3-dimethylpiperazine-1-carboxylateor tert-butyl(2R,3S)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,3-dimethylpiperazine-1-carboxylate(Intermediate CE) (248 mg, 357 μmol) at RT and reactants were stirred atRT for 1 hour. The RM was diluted with DCM/NaHCO₃, extracted twice withDCM and the combined organic extracts were washed with brine, dried overNa₂SO₄ and concentrated to give the title compound as a white solid.

LC-MS: Rt=0.81 min; MS m/z [M+H]⁺ 594.5/596.5, m/z [M−H]⁻ 592.3/594.3;UPLC-MS 1

Intermediate CL:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-((2S,3R)-2,3-dimethylpiperazin-1-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideorN-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-((2R,3S)-2,3-dimethylpiperazin-1-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

HCl 4N in 1,4-dioxane (1.00 mL, 4.00 mmol) was added to tert-butyl(2R,3S)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,3-dimethylpiperazine-1-carboxylateor tert-butyl(2S,3R)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,3-dimethylpiperazine-1-carboxylate(Intermediate CG) (258 mg, 357 μmol) at RT and reactants were stirred atRT for 15 minutes. The RM was diluted with DCM/NaHCO₃, extracted twicewith DCM and the combined organic extracts were washed with brine, driedover Na₂SO₄ and concentrated to give the title compound as a reddishsolid.

LC-MS: Rt=0.79 min; MS m/z [M+H]⁺ 594.3/596.2, m/z [M−H]⁻ 592.2/594.3;UPLC-MS 1

Intermediate AF: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylateStep 1: tert-butyl4-(3-amino-6-ethyl-2-imino-4-oxo-1,2,3,4-tetrahydropyrimidin-5-yl)piperazine-1-carboxylate

Hydrazinecarboximidamide.HCl (35.0 g, 317 mmol) was mixed with EtOH (400mL), followed by aqueous tetrabutylammonium hydroxide solution, 40 wt %in water (206 g, 318 mmol). The mixture was stirred at 55° C. for 80minutes. Tert-butyl4-(1-methoxy-1,3-dioxopentan-2-yl)piperazine-1-carboxylate (IntermediateEH) (50.0 g, 159 mmol) was added. The mixture was stirred at reflux for6 hours, and then it was cooled to RT. The solvent was removed undervacuum until % of the solvent volume remained. The resulting suspensionwas stirred for 1-2 hours, then filtered. The cake was dried undervacuum to give the title compound as a white solid.

MS m/z [M+H]⁺ 339.2

Step 2: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

A solution of 3,6-dihydro-2H-pyran-4-carbaldehyde (Intermediate DK)(20.0 g, 178 mmol) in NMP and tert-butyl4-(3-amino-6-ethyl-2-imino-4-oxo-1,2,3,4-tetrahydropyrimidin-5-yl)piperazine-1-carboxylate(52.9 g, 149 mmol) in NMP (400 mL), followed by FeCl₃ (48 g, 297 mmol).The dark solution was heated to 50° C. open to air and stirred for 48hours. The dark RM was cooled to RT. Water (1.2 L) was added slowly(exothermic). The suspension was filtered and washed with water (400mL). The resulting wet cake was added to acetone (400 mL), stirred at RTfor 4 hours. The suspension was filtered and washed with acetone (100mL). The cake was added to EtOH (400 mL) and heated to 70° C., stirredfor 4 hours. Then the mixture was cooled to RT, filtered and washed withEtOH (100 mL) to give the title compound as brown solid.

MS m/z [M+H]⁺ 430.2

Intermediate AK:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: tert-butyl4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

A solution of tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate AF) (50.0 g, 116 mmol),2-chloro-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (IntermediateDL step 1) (56.9 g, 209 mmol), DIPEA (60.9 mL, 348 mmol) in NMP (500 mL)was stirred at 50° C. for 16 hours. Water (1.5 L) was added to the RMand it was stirred for 1.5 hours, filtered and washed with water (600mL). The resulting brown solid was added to EtOH (500 mL), stirred at RTfor 4 hours, then it was filtered and washed with EtOH (150 mL). Thecake was added to THF (1 L) and heated to 65° C., heptane (1 L) wasadded and it was stirred for 30 minutes. Then the mixture was cooled toRT. The suspension was filtered and washed with heptane (100 mL). Thewet cake was dried to give the title compound as a grey solid.

MS m/z [M+H]⁺ 666.4

Step 2:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

Tert-butyl4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(3.10 g, 3.96 mmol) was mixed with DCM (225 mL) and HCl 2M in EtOAc (135mL, 270 mmol). The suspension was stirred at RT for 6 hours. Then addedDCM (305 mL) and HCl 2M in EtOAc (185 mL, 370 mmol), followed by solidtert-butyl4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(57.4 g, 73.2 mmol). The RM was stirred at RT for 6 hours. Thesuspension was filtered and rinsed twice with premixed DCM:EtOAc (1:1,220 mL). The cake was collected and dried under vacuum at RT for 4hours. The dry cake was mixed with a premixed solution of Et₃N (16.1 mL,116 mmol) in ACN:water (1:1, 640 mL). The suspension was stirred at RTfor 1 hour. Then water (1.6 L) was added over 2 hours, then stirred for6 hours. The suspension was filtered and rinsed twice with ACN:water(1:4, 270 mL). The wet cake was dried under vacuum at 55° C. for 6 hoursto give the title compound as a light brown solid.

MS m/z [M+H]⁺ 566.2

Intermediate CM: tert-butyl5-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylateStep 1: 2-bromo-5-ethyl-[1,2,4]triazolo[1,5-a]pyrimidin-7(4H)-one

3-Bromo-1H-1,2,4-triazol-5-amine (Intermediate ER) (20.0 g, 123 mmol)and ethyl 3-oxopentanoate (22.1 g, 153 mmol) were mixed in AcOH (70 mL)and stirred at 80° C. for 19 hours. The RM was cooled to RT, then it wasstirred at 0° C. for 1 hour. The resulting suspension was filtered andwashed with a small amount of EtOH. The obtained solid was dried underHV to give the title compound as a white solid. The filtrate wasconcentrated and crystallized from EtOH to afford a second batch ofproduct, which was dried under HV to give the title compound as a beigesolid.

LC-MS: Rt=0.23 min; MS m/z [M+H]⁺ 243.0/245.0, m/z [M−H]⁻ 241.1/243.1;UPLC-MS 1

Step 2:2-(2-bromo-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

2-Bromo-5-ethyl-[1,2,4]triazolo[1,5-a]pyrimidin-7(4H)-one (7.00 g, 28.8mmol) and, N-(2-chloro-4-(trifluoromethyl)phenyl)-2-iodoacetamide(Intermediate DL) (11.8 g, 31.7 mmol) were mixed in 1,4-dioxane (170 mL)at RT under argon. DIPEA (15.1 mL, 86.0 mmol) was added and the RM wasstirred at 80° C. for 4.25 hours. The RM was cooled to RT and extractedwith EtOAc (3×300 mL), water (2×75 mL) and brine (2×75 mL). The aqueouslayer was a suspension, which was filtered. The solid was dissolved inDCM/MeOH and combined with the organic layer. The combined organiclayers were dried through a phase separator and concentrated underreduced pressure. The resulting bright brown solid was suspended in Et₂Oand sonicated for 30 minutes, then it was stirred at reflux for 3 hours.The suspension was filtered, washed with Et₂O and dried under HV to givethe title compound as a beige solid.

LC-MS: Rt=0.99 min; MS m/z [M+H]⁺ 478.1/480.1/482.1, m/z [M−H]⁻476.1/478.1/480.1; UPLC-MS 1

Step 3:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

2-(2-Bromo-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(14.0 g, 29.2 mmol), K₃PO₄ (18.6 g, 88.0 mmol) and Pd(dppf)Cl₂.DCM (1.19g, 1.46 mmol) were mixed at RT, evacuated and purged with argon severaltimes. 1,4-Dioxane (300 mL) and water (150 mL) were added and the RM wasevacuated and purged with argon several times.2-(3,6-Dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(7.99 g, 38.0 mmol) was added and the RM was stirred at 80° C. for 1hour. The RM was concentrated under reduced pressure. The residue wasextracted with 10% MeOH in DCM (4×400 mL) and water (2×150 mL). Thecombined organic layers were dried through a phase separator and thefiltrate was concentrated to half volume under reduced pressure.ISOLUTE® Si-Thiol (15.0 g) was added and the suspension was stirred for30 minutes, the solid was filtered and washed sequentially withDCM/MeOH, 1,4-dioxane, warm MeOH and warm EtOH. The filtrates wereconcentrated under reduced pressure to give 2 batches of the titlecompound as a white solid and as a brown solid.

LC-MS: Rt=0.96 min; MS m/z [M+H]⁺ 482.2/484.2, m/z [M−H]⁻ 480.3/482.3;UPLC-MS 1

Step 4:2-(6-bromo-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide(1.20 g, 2.49 mmol) and NBS (488 mg, 2.74 mmol) were suspended in ACN(50 mL) and the RM was stirred at RT for 19.5 hours. The RM was quenchedwith water and extracted with DCM (3×150 mL). The organic layer wasdried through a phase separator and concentrated under reduced pressure.The crude product was adsorbed onto Isolute and purified by columnchromatography (RediSep Column: Silica 40 g, eluent DCM:DCM/MeOH (8/2)100:0 to 50:50). The product containing fractions were combined,concentrated and dried under HV to give a beige solid. The beige solidwas adsorbed onto Isolute and purified by column chromatography (RediSepColumn: Silica 40 g, eluent heptane:EtOAc 100:0 to 0:100, thenDCM:DCM/MeOH (8/2) 50:50 to 0:100). The product containing fractionswere combined, concentrated and dried under HV to give the titlecompound as a pale beige solid.

LC-MS: Rt=1.07 min; MS m/z [M+H]⁺ 560.1/562.1/564.0, m/z [M−H]⁻558.1/560.1/562.0; UPLC-MS 1

Step 5: tert-butyl5-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylate

2-(6-Bromo-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(400 mg, 713 μmol), tert-butyl2,5-diazabicyclo[4.2.0]octane-2-carboxylate (1.51 g, 7.13 mmol) andAgBF₄ (140 mg, 713 μmol) were mixed in DMSO (6 mL) and the RM wasstirred at 120° C. in a sealed MW vial for 23 hours. The RM was cooledto RT. The RM was extracted with DCM (4×60 mL), aq NaHCO₃ (2×30 mL) andbrine (30 mL). The organic layer was dried through a phase separator andconcentrated under reduced pressure to give a brown oil. The crudeproduct was adsorbed onto Isolute and purified by column chromatography(RediSep Column: Silica 40 g, eluent heptane:EtOAc 100:0 to 0:100). Theproduct containing fractions were combined, concentrated and dried underHV to give a bright beige solid.

Chiral separation of rac-tert-butyl5-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylate:

Preparative chiral SFC (instrument: Sepiatec Prep SFC100; column: OVEN3Chiralpak IB-N 250×30 mm, 5 μm; eluent: A: 35% [IPA+0.1% NH₃] B:65%scCO2; flow rate: 80.0 mL/min; detection: 210 nm; back pressure: 120bar; injection volume: 1 mL; Gradient: isocratic)

Intermediate CN: tert-butyl(1R,6R)-5-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylate

First Eluting Stereoisomer

Chiral HPLC (C-HPLC 10): Rt=2.1 min, 97% ee

LC-MS: Rt=1.38 min; MS m/z [M+H]⁺ 692.3/694.3, m/z [M−H]⁻ 690.4/692.4;UPLC-MS 1

Intermediate CO: tert-butyl(1S,6S)-5-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylate

Second Eluting Stereoisomer

Chiral HPLC (C-HPLC 10): Rt=2.6 min, 95% ee

LC-MS: Rt=1.38 min; MS m/z [M+H]⁺ 692.3/694.2, m/z [M−H]⁻ 690.3/692.3;UPLC-MS 1

Intermediate CP:2-(6-((1R,6R)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

Tert-butyl(1R,6R)-5-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylate(Intermediate CN) (first eluting stereoisomer—150 mg, 217 μmol) wasdissolved in DCM (3.1 mL) at RT. TFA (334 μL, 4.33 mmol) was added andthe RM was stirred at RT for 2 hours. The RM was concentrated underreduced pressure and dried under HV to give the title compoundtrifluoroacetate salt as a bright brown solid.

LC-MS: Rt=0.78 min; MS m/z [M+H]⁺ 592.3/594.3, m/z [M−H]⁻ 590.2/592.2;UPLC-MS 1

Intermediate CQ:2-(6-((1S,6S)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

Tert-butyl(1S,6S)-5-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylate(Intermediate CO) (second eluting stereoisomer—128 mg, 185 μmol) wasdissolved in DCM (2.6 mL) at RT. TFA (285 μL, 3.70 mmol) was added andthe RM was stirred at RT for 1.25 hours. The RM was concentrated underreduced pressure and dried under HV to give the title compoundtrifluoroacetate salt as a bright brown solid.

LC-MS: Rt=0.79 min; MS m/z [M+H]⁺ 592.3/594.2, m/z [M−H]⁻ 590.3/592.2;UPLC-MS 1

Intermediate CR: tert-butyl4-(5-methyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-6-(piperazin-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)-3,6-dihydropyridine-1(2H)-carboxylateStep 1: tert-butyl4-(2-bromo-5-methyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate BM) (4.00 g, 9.68 mmol) and2-bromo-N-(4-(trifluoromethyl)phenyl)acetamide (3.28 g, 11.6 mmol) weremixed with DMF (25 mL) at RT. DIPEA (5.07 mL, 29.0 mmol) was added andthe RM was stirred at 85° C. for 2.3 hours. The RM was cooled to RT andwater (50 mL) was added slowly. The suspension was stirred at RT. Thenit was cooled to 0° C. and filtered. The cake was washed with water anddried to give the title compound.

LC-MS: Rt=1.24 min; MS m/z [M+H−Boc]⁺ 514.1/516.1, m/z [M−H]⁻612.2/614.2; UPLC-MS 8

Step 2:2-(2-bromo-5-methyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

Tert-butyl4-(2-bromo-5-methyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(5.67 g, 9.23 mmol) was mixed with DCM (25 mL). TFA (14.2 mL, 185 mmol)was added and the RM was stirred at RT for 40 minutes. The RM wasconcentrated under reduced pressure, redissolved in DCM and concentratedunder reduced pressure again. This was performed three times. Theresulting oil was dried under HV, extracted three times with EtOAc, oncewith aq sat NaHCO₃ and twice with brine. The combined organic layerswere dried through a phase separator and concentrated under reducedpressure. The resulting brown solid was suspended in Et₂O and sonicated.Then it was filtered and washed with Et₂O to give the title compound asa bright brown solid.

LC-MS: Rt=0.78 min; MS m/z [M+H]⁺ 514.1/516.1, m/z [M−H]⁻ 512.0/514.0;UPLC-MS 8

Step 3: tert-butyl4-(5-methyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-6-(piperazin-1-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate

2-(2-Bromo-5-methyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide(500 mg, 924 μmol), tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate(376 mg, 1.15 mmol), XPhos Pd G3 (39.1 mg, 46.0 μmol), 1,4-dioxane (4.2mL) and K₃PO₄ 1M in water (2.77 mL, 2.77 mmol) were mixed. The RM wasvacuumed and purged with argon several times. Then it was stirred at 90°C. for 1.5 hours. The RM was cooled to RT and the 1,4-dioxane wasremoved under reduced pressure. The residue was extracted with EtOAc(3×50 mL), water (2×10 mL) and brine (2×5 mL). The organic layer wasdried through a phase separator and concentrated under reduced pressure.The residue was dissolved in DCM/MeOH and ISOLUTE® Si-Thiol (750 mg) wasadded. The suspension was stirred for 20 minutes, then it was filteredand the cake was washed with DCM and MeOH. The filtrate was concentratedunder reduced pressure. The residue was suspended in Et₂O, sonicated andfiltered. The solid was dried under HV to give the title compound as abeige solid.

LC-MS: Rt=0.99 min; MS m/z [M+H]⁺ 617.3, m/z [M−H]⁻ 615.2; UPLC-MS 4

Intermediate CS:(S)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoro-3,4-dihydro-2H-pyran-6-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamideStep 1: methyl6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidine-2-carboxylate

Tert-butyl4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate X) (6.00 g, 14.0 mmol), Pd(dppf)Cl₂.DCM (573 mg, 702μmol), 1,1′-ferrocenediyl-bis(diphenylphosphin (389 mg, 702 μmol) andMeOH (140 mL) were mixed. Then Et₃N (3.91 mL, 28.1 mmol) was added. TheRM was set under CO gas (15 bar) and stirred at 100° C. for 22 hours.The red yellow fine suspension was filtered. The orange filter cake waswashed with DCM and the filtrate was concentrated under reducedpressure. The crude product was purified by column chromatography(Silica gel column: Silica 120 g, eluent DCM:MeOH 100:0 to 90:10). Theproduct containing fractions were combined and concentrated underreduced pressure (no separation). The crude product was purified bycolumn chromatography (Silica gel column: Silica 120 g, eluent TBME:MeOH100:0 to 60:40). The product containing fractions were combined andconcentrated under reduced pressure to give the title compound.

LC-MS: Rt=0.84 min; MS m/z [M+H]⁺ 407.5, m/z [M−H]⁻ 405.5; UPLC-MS 1

Step 2: methyl6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-5-ethyl-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidine-2-carboxylate

A 300 mL 4 neck reaction flask with mechanical stirrer, internalthermometer, dropping funnel and nitrogen inlet was charged with methyl6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidine-2-carboxylate(7.08 g, 13.2 mmol) (first as a solid the rest was added with thesolvent). THF (100 mL) and DMF (20 mL) were added and the brownsuspension was cooled in an ice bath under nitrogen. After 10 minutessodium hydride (980 mg, 24.5 mmol) was added in portions. After another30 minutes in the ice bath a solution of(2-(chloromethoxy)ethyl)trimethylsilane (4.34 mL, 24.5 mmol) in THE (20mL) was added within 5 minutes. The ice bath was removed and the mixturewas stirred at RT for 2 hours. The RM was cooled in an ice bath andstirred overnight. Additional sodium hydride (490 mg, 12.3 mmol) wasadded at RT and the RM was stirred for 4.5 hours.(2-(chloromethoxy)ethyl)trimethylsilane (2.17 mL, 12.3 mmol) was addedand the RM was stirred for 40 minutes. The RM was slowly poured into anaq solution of water/ice (1:1) containing citric acid (9.67 g, 50.3mmol). TBME (200 mL) was added and the phases were separated. A finepurple slurry remained between the phases that was poured into theaqueous phase, which was extracted with TBME (3×200 mL). The combinedorganic phases were washed with brine (100 mL) and dried over MgSO₄ andconcentrated under reduced pressure to give a purple, viscous oil. Thecrude product was purified by column chromatography (Silica gel column:Silica 120 g, eluent heptane:EtOAc). The product containing fractionswere combined and concentrated under reduced pressure to give the titlecompound as an isomeric mixture (SEM group also attached to 5-memberedring) as a white foam.

LC-MS: Rt=1.37/1.49 min; MS m/z [M+H]⁺ 537.3; UPLC-MS 1

Step 3:6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-5-ethyl-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidine-2-carboxylicacid

Methyl6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-5-ethyl-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidine-2-carboxylate(3.34 g, 6.09 mmol) was dissolved in THE (1.5 mL) and cooled in an icebath. After 10 minutes, a solution of sodium hydroxide (375 mg, 9.37mmol) in water (1.5 mL) was added. The mixture was stirred in the icebath for 1.2 hours. The ice bath was removed and the RM was stirred atRT for 2 hours. Addition of a solution of sodium hydroxide (621 mg, 15.5mmol) in water (2 mL). The RM was stirred at RT for 3.75 hours. Additionof HCl 1M (25.0 mL, 25.0 mmol) until pH=7. The solvent was evaporated at40° C. in vacuo, a gum was formed that was dissolved in EtOAc (50 mL).After phase separation the aqueous phase was extracted with EtOAc (3×20mL). The combined organic phases were dried over MgSO₄, filtered andconcentrated under reduced pressure to give an isomeric mixture of thetitle compound as a white powder.

LC-MS: Rt=1.18/1.22 min; MS m/z [M+H]⁺ 523.3, m/z [M−H]⁻ 521.4; UPLC-MS1

Step 4: tert-butyl4-(5-ethyl-2-(methoxy(methyl)carbamoyl)-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

A solution of6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-5-ethyl-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidine-2-carboxylicacid (2.93 g, 5.61 mmol), Et₃N (1.88 mL, 13.5 mmol) andN,O-dimethylhydroxylamine hydrochloride (1.09 g, 11.2 mmol) in dry DCM(50 mL) was cooled in an ice bath for 10 minutes, then slowly T₃P 50% inEtOAc (4.28 g, 6.73 mmol) was added. The ice bath was removed and the RMwas stirred at RT for 3.5 hours. The RM was cooled in an ice bath andquenched with sat. NaHCO₃ (40 mL). After phase separation, the aqueousphase was extracted with DCM (2×10 mL). The combined DCM phases werewashed with aqueous citric acid followed by brine. The organic phase wasdried over MgSO₄, filtered and concentrated to give a pale brown foam.The crude product was purified by column chromatography (Silica gelcolumn: Silica 120 g, eluent heptane:EtOAc 70:30 to 0:100). The productcontaining fractions were combined and concentrated under reducedpressure to give the title compound as an isomeric mixture.

LC-MS: Rt=1.34/1.49 min; MS m/z [M+H]⁺ 566.5/567.5; UPLC-MS 1

Step 5: tert-butyl(S)-4-(2-(5-(benzyloxy)-4-fluoropentanoyl)-5-ethyl-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

(S)-((2-fluoro-4-iodobutoxy)methyl)benzene (Intermediate EG) (380 mg,1.23 mmol) and tert-butyl4-(5-ethyl-2-(methoxy(methyl)carbamoyl)-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(250 mg, 411 μmol) was dissolved in THE (19 mL) and pentane (19 mL). Thecolorless clear RM was cooled in a liquid nitrogen/EtOH bath to ca −100°C. Then tert-butyllithium 1.7M in pentane (1.45 mL, 2.47 mmol) was addeddropwise within 10 minutes (using a syringe pump with a steel cannulawhich is immersed in the solution 145 μL/min). The RM was stirred at ca−100° C. for 60 minutes. The −100° C. cold RM was poured rapidly into avigorously stirred mixture of 10% citric acid solution (50 mL) and TBME(50 mL) and stirred for 5 minutes. The phases were separated. Theorganic phase was extracted once with brine (50 mL). The aqueous phaseswas extracted with TBME (2×25 mL). The combined organic phases weredried over anhydrous Na₂SO₄, filtered and concentrated in vacuo at 45°C. to give a yellow resin. The crude product was purified by columnchromatography (Silica gel column: Silica 40 g, eluent heptane:EtOAc90:10 to 0:100). The product containing fractions were combined andconcentrated under reduced pressure to give the title compound as acolourless resin.

LC-MS: Rt=1.62 min; MS m/z [M+H−Boc]⁺ 587.5, m/z [M+H]⁺ 687.6; UPLC-MS 1

Step 6: tert-butyl(S)-4-(5-ethyl-2-(4-fluoro-5-hydroxypentanoyl)-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To a colorless solution of tert-butyl(S)-4-(2-(5-(benzyloxy)-4-fluoropentanoyl)-5-ethyl-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(170 mg, 247 μmol) in MeOH (8 mL) was added 10% palladium on carbon(26.3 mg, 25.0 μmol). The RM was hydrogenated at RT for 45 minutes. Tothe RM was added further palladium on carbon 10% (52.7 mg, 49.0 μmol)and hydrogenated at RT overnight. The RM was filtered through asyringe-filter, washed with MeOH and concentrated in vacuo at 45° C. togive the title compound as colourless resin.

LC-MS: Rt=1.34 min; MS m/z [M+H−Boc]⁺ 497.4; UPLC-MS 1

Step 7: tert-butyl(S)-4-(5-ethyl-2-(3-fluoro-3,4-dihydro-2H-pyran-6-yl)-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To a solution of tert-butyl(S)-4-(5-ethyl-2-(4-fluoro-5-hydroxypentanoyl)-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(127 mg, 160 μmol) in toluene (6 mL), p-toluenesulfonic acid monohydrate(27.3 mg, 144 μmol) and Molecular sieves 3A (800 mg, 160 μmol) wereadded. The RM was heated to 45° C. and stirred for 20 hours. The RM wasextracted with a pH 6.0 buffer (20 mL) and DCM (30 mL). The aqueousphase was extracted with DCM (3×20 mL). The combined organic phases weredried over anhydrous Na₂SO₄, filtered and concentrated in vacuo at 45°C. to give a pale yellow resin. The crude product was purified by columnchromatography (Silica gel column: Silica 40 g, eluent DCM:DCM/MeOH(9/1) 100:0 to 20:80). The product containing fractions were combinedand concentrated under reduced pressure to give the title compound as acolourless resin as well as 2 batches of tert-butyl(S)-4-(5-ethyl-2-(3-fluoro-3,4-dihydro-2H-pyran-6-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate.

LC-MS: Rt=1.43 min; MS m/z [M+H]⁺ 479.4; UPLC-MS 1

Step 8: tert-butyl(S)-4-(5-ethyl-2-(3-fluoro-3,4-dihydro-2H-pyran-6-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To a colorless solution of tert-butyl(S)-4-(5-ethyl-2-(3-fluoro-3,4-dihydro-2H-pyran-6-yl)-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(19.0 mg, 18.0 μmol) in THF (1 mL) was added tetrabutylammonium fluoride1M in THF (90.0 μL, 90.0 μmol). The RM was heated to 40° C. and stirredovernight. The RM was evaporated in vacuo at 45° C. to give a pale brownoil. The crude product was purified by column chromatography (Silica gelcolumn: Silica 40 g, eluent heptane:EtOAc 80:20 to 0:100, thenDCM:DCM/MeOH (9/1) 100:0 to 0:100). The product containing fractionswere combined and concentrated under reduced pressure to give the titlecompound as a white solid.

LC-MS: Rt=0.88 min; MS m/z [M+H]⁺ 449.6, m/z [M−H]⁻ 447.5; UPLC-MS 1

Step 9: tert-butyl(S)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-(3-fluoro-3,4-dihydro-2H-pyran-6-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

To a pale yellow solution of tert-butyl(S)-4-(5-ethyl-2-(3-fluoro-3,4-dihydro-2H-pyran-6-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(35.0 mg, 78.0 μmol) andN-(2-chloro-4-(trifluoromethyl)phenyl)-2-iodoacetamide (Intermediate DL)(31.2 mg, 86.0 μmol) in 1,4-dioxane (5 mL) was added DIPEA (41.0 μL, 234μmol). The RM was heated to 80° C. and stirred for 11 hours, then it wasleft to stand at RT overnight. DIPEA (41.0 μL, 234 μmol) was added againand the RM was stirred at 80° C. for 6 hours. The RM was concentrated invacuo at 45° C. to give a pale brown resin (103 mg). The crude productwas purified by column chromatography (Silica gel column: 40 g SNAPcartridge (Biotage), eluent heptane:EtOAc 80:20 to 0:100). The productcontaining fractions were combined and concentrated under reducedpressure to give the title compound as a pale brown residue.

LC-MS: Rt=1.31 min; MS m/z [M+H−Boc]⁺ 584.3/586.3, m/z [M−H]⁻682.4/684.4; UPLC-MS 1

Step 10:(S)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoro-3,4-dihydro-2H-pyran-6-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

To a solution of tert-butyl(S)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-(3-fluoro-3,4-dihydro-2H-pyran-6-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(42.0 mg, 53.4 μmol) in DCM (250 μL), TFA (247 μL, 3.21 mmol) was addedand the RM was stirred at RT for 1 hour. The RM was concentrated invacuo at 45° C. The residue was evaporated three times with DCM to givea yellow resin. The residue was diluted with DCM (10 mL) and extractedonce with aq sat NaHCO₃ (10 mL). The aqueous phase was extracted withDCM (3×10 mL). The combined organic phases were dried over anhydrousNa₂SO₄, filtered and concentrated in vacuo at 45° C. to give a paleyellow resin. The crude product was purified by column chromatography(Silica gel column: 40 g SNAP cartridge (Biotage), eluentDCM:DCM/MeOH/NH₃ (85/15/1) 100:0 to 0:100). The product containingfractions were combined and concentrated under reduced pressure to givea colourless resin. The crude product was purified by columnchromatography (Silica gel column: 24 g SNAP cartridge (Biotage), eluentDCM:DCM/MeOH/NH₃ (85/15/1) 100:0 to 0:100). The product containingfractions were combined and concentrated under reduced pressure to givethe title compound as a pale yellow solid.

LC-MS: Rt=0.78 min; MS m/z [M+H]⁺ 584.3/586.3, m/z [M−H]⁻ 582.3/584.3;UPLC-MS 1

Preparation of Early Intermediates

Intermediate CT: perfluorophenyl 3-hydroxypicolinate

3-Hydroxypicolinic acid (8.00 g, 57.5 mmol) was suspended in DCM (50mL). 2,3,4,5,6-pentafluorophenol (9.53 g, 51.8 mmol) anddiisopropylmethanediimine (10.8 mL, 69.0 mmol) were added. The RM wasstirred at RT for 12 hours, concentrated under reduced pressure and thecrude product was purified by column chromatography (Silica gel column:Silica 40 g, eluent hexane:EtOAc 100:0 to 95:5). The product containingfractions were combined and concentrated under reduced pressure to givethe title compound.

HPLC: Rt=5.97 min; HPLC 1

Intermediate CU: 3-(benzyloxy)-4-fluoropicolinic acid Step 1:2-chloro-4-fluoro-3-((4-methoxybenzyl)oxy)pyridine

2-Chloro-4-fluoropyridin-3-ol (3.00 g, 19.3 mml) and K₂CO₃ (4.00 g, 29.0mmol) were dissolved in DMF (55 mL). The RM was flushed with argon,1-(chloromethyl)-4-methoxybenzene (2.15 mL, 21.3 mmol) was added and theRM was stirred at 60° C. for 4 hours. EtOAc (100 mL) was added and itwas washed with water (2×50 mL) and brine (50 mL), dried over Na₂SO₄,filtered and evaporated. The crude product was purified by columnchromatography (RediSep Column: Silica 80 g, eluent heptane/EtOAc 100:0to 75:25). The product containing fractions were combined, concentratedunder reduced pressure and dried under HV to give the title compound.

LC-MS: Rt=1.08 min; MS m/z [M+H]⁺ 268.0/270.0; UPLC-MS 1

Step 2: methyl 4-fluoro-3-hydroxypicolinate

To a stirred and degassed solution of2-chloro-4-fluoro-3-((4-methoxybenzyl)oxy)pyridine (3.44 g, 12.8 mmol)in Et₃N (3.58 mL, 25.7 mmol) and MeOH (50 mL) was added Pd(dppf)Cl₂.DCM(524 mg, 642 μmol) and the RM was stirred at 50° C. under 10 bars of COuntil completion. The RM was filtered through celite and concentratedunder reduced pressure. The crude product was purified by columnchromatography (RediSep Column: Silica 40 g, eluent heptane:EtOAc 100:0to 0:100). The product containing fractions were combined andconcentrated under reduced pressure to give the title compound as awhite solid.

LC-MS: Rt=0.37 min; MS m/z [M+H]⁺ 172.1; UPLC-MS 1

Step 3: methyl 3-(benzyloxy)-4-fluoropicolinate

To a stirred solution of methyl 4-fluoro-3-hydroxypicolinate (500 mg,2.92 mmol) in DMF (5 mL) were added K₂CO₃ (606 mg, 4.38 mmol) and(bromomethyl)benzene (382 μL, 3.21 mmol) and reactants were stirred atRT for 14 hours. The RM was diluted with EtOAc/water, extracted withEtOAc and the organic layer was washed with water and brine, dried overNa₂SO₄ and concentrated under reduced pressure. The crude product waspurified by column chromatography (RediSep Column: Silica 24 g, eluentheptane:EtOAc 100:0 to 50:50). The product containing fractions werecombined and concentrated under reduced pressure to afford the titlecompound as a yellow oil.

LC-MS: Rt=0.89 min; MS m/z [M+H]⁺ 262.0; UPLC-MS 1

Step 4: 3-(benzyloxy)-4-fluoropicolinic acid

To a stirred solution of methyl 3-(benzyloxy)-4-fluoropicolinate (750mg, 2.87 mmol) in THF (5 mL) and MeOH (5 mL) was added NaOH 2N in water(5.00 mL, 10.0 mmol) at RT and the RM was stirred at RT for 30 minutes.THF and MeOH were removed under reduced pressure, then the resultingaqueous residue was acidified to pH 3 with 2N HCl and extracted threetimes with DCM. The combined organic layers were dried over Na₂SO₄ andconcentrated under reduced pressure to give the title compound as awhite solid.

LC-MS: Rt=0.58 min; MS m/z [M+H]⁺ 248.1, m/z [M−H]⁻ 246.1; UPLC-MS 1

Intermediate CV: 3-hydroxypicolinoyl chloride

3-Hydroxypicolinic acid (12.0 g, 82.0 mmol) and sulfurous dichloride(121 mL, 1.64 mol) were mixed under argon at RT. The flask was equippedwith a condenser and a drying tube (CaCl₂) and the suspension wasstirred at RT for 5 days. Then it was filtered and the cake was washedseveral times with TBME. The solid was dried at 30° C. under HV andstored under argon.

Intermediate CW: 5-methoxy-6-methylpyrimidine-4-carboxylic acid Step 1:4-chloro-5-methoxy-6-methylpyrimidine

4,6-Dichloro-5-methoxypyrimidine (75.0 g, 419 mmol), methylboronic acid(26.4 g, 441 mmol) and K₃PO₄ (222 g, 1.05 mol), Pd(dppf)Cl₂.DCM (20.1 g,24.6 mmol) were suspended in DME (300 mL). The RM was flushed with argonthree times. The RM was heated at 85° C. for 18 hours. The RM wasfiltered through celite and the solution was concentrated under reducedpressure. The residue was dissolved in EtOAc (500 mL) and washed with aqsat NaHCO₃ (3×800 mL). The organic layer was collected, concentratedunder reduced pressure and dried. The crude product was purified bycolumn chromatography (silica column 60 mm×475 mm×50 mm, eluentheptane:EtOAc 50:1 to 20:1) to give the title compound as a solid.

HPLC: Rt=2.93 min; HPLC 4

Step 2: methyl 5-methoxy-6-methylpyrimidine-4-carboxylate

4-Chloro-5-methoxy-6-methylpyrimidine (24.0 g, 129 mmol),Pd(dppf)Cl₂.DCM (6.27 g, 7.68 mmol) and Et₃N (35.5 mL, 256 mmol) weredissolved in DCM (600 mL). The RM was purged with CO for 3 times. The RMwas heated at 60° C. under CO (0.3 PMa) for 18 hours. The RM wasfiltered through celite and the solution was concentrated under reducedpressure. The residue was purified by column chromatography (silicacolumn 60 mm×475 mm×50 mm, eluent heptane:EtOAc 50: 1 to 30:1) to givethe title compound as a solid.

HPLC: Rt=2.55 min; HPLC 4

Step 3: 5-methoxy-6-methylpyrimidine-4-carboxylic acid

To a solution of methyl 5-methoxy-6-methylpyrimidine-4-carboxylate (15.0g, 76.0 mmol) in THF (140 mL) and water (70 mL) was added LiOH.H₂O (3.50g, 83.0 mmol). The RM was stirred at RT for 16 hours. The RM wasconcentrated under reduced pressure to give the title compound as alithium salt.

HPLC: Rt=2.10 min, HPLC 3

Intermediate CX: 3-(benzyloxy)-4-fluoro-6-methylpicolinic acid Step 1:2-bromo-6-methyl-4-nitropyridin-3-ol

2-Bromo-6-methylpyridin-3-ol (15.0 g, 80.0 mmol) was mixed with H₂SO₄(44.8 mL, 798 mmol) at 0° C. HNO₃ (66%) (6.03 mL, 88.0 mmol) was addeddropwise within 12 minutes. The orange solution was stirred at 0° C. for1 hour, then at RT for 3 hours. HNO₃ (66%) (1.10 mL, 16.0 mmol) wasadded and the reaction was stirred at RT for 3 hours. The RM was pouredwhile vigorous stirring onto ice. The mixture was extracted with EtOAc(3×600 mL). The organic layer was washed with water (2×75 mL) and brine(2×75 mL), dried over Na₂SO₄, filtered and concentrated under reducedpressure. The residue was adsorbed onto Isolute and purified by columnchromatography (RediSep Column: Silica 40 g, eluent DCM:DCM/MeOH (8/2)100:0 to 0:100). The product containing fractions were combined andconcentrated under reduced pressure to give the title compound as ayellow solid.

LC-MS: Rt=0.53 min; MS m/z [M−H]⁻ 231.0/233.0; LC-MS 1

Step 2: 3-(benzyloxy)-2-bromo-6-methyl-4-nitropyridine

2-Bromo-6-methyl-4-nitropyridin-3-ol (10.4 g, 44.5 mmol) and K₂CO₃ (18.5g, 134 mmol) were dissolved in DMF (110 mL) at RT under argon.(bromomethyl)benzene (11.7 g, 66.8 mmol) was added and the RM wasstirred at 45° C. for 17 hours. The RM was cooled to RT, filtered andthe cake was washed with ACN. The filtrate was concentrated underreduced pressure, the residue was adsorbed onto Isolute and purified bycolumn chromatography (RediSep Column: Silica 220 g, eluent DCM). Theproduct containing fractions were combined and concentrated underreduced pressure to give the title compound as an orange oil.

LC-MS: Rt=1.25 min; MS m/z [M+H]⁺ 323.1/325.1; LC-MS 1

Step 3: 3-(benzyloxy)-2-bromo-4-fluoro-6-methylpyridine

3-(Benzyloxy)-2-bromo-6-methyl-4-nitropyridine (9.30 g, 28.8 mmol) wasdissolved in DMF (150 mL). The flask was vacuumed and purged with argonseveral times. TBAF 1M in THF (58.0 mL, 58.0 mmol) was added and the RMwas stirred at RT for 45 minutes. TBAF 1M in THF (20.0 mL, 20.0 mmol)was added and the RM was stirred at RT for 1.75 hours. The RM wasquenched with water (250 mL) and extracted with EtOAC (3×400 mL). Theorganic layer was washed with water (200 mL) and brine (2×150 mL), driedover Na₂SO₄ and filtered. The filtrate was concentrated under reducedpressure. The crude product was purified by column chromatography(RediSep Column: Silica 220 g, eluent DCM). The product containingfractions were combined and concentrated under reduced pressure. Theresidue was purified by column chromatography (RediSep Column: Silica120 g, eluent DCM). The product containing fractions were combined andconcentrated under reduced pressure to give the title compound as abeige solid.

LC-MS: Rt=1.23 min; MS m/z [M+H]⁺ 296.1/298.1; LC-MS 1

Step 4: ethyl 3-(benzyloxy)-4-fluoro-6-methylpicolinate

A solution of 3-(benzyloxy)-2-bromo-4-fluoro-6-methylpyridine (4.93 g,16.7 mmol), Pd(dppf)Cl₂.DCM (680 mg, 832 μmol) and Et₃N (7.03 mL, 49.9mmol) in EtOH (520 mL) was stirred at 80° C. under 18 bar of CO for 20hours. SiliaMetS®Thiol was added, the mixture was stirred at 40° C. for1 hour, filtered and the cake was washed with DCM. The filtrate wasconcentrated under reduced pressure, the crude product was suspended inDCM, filtered, washed with Et₂O and purified by column chromatography(RediSep Column: Silica 120 g, eluent DCM:DCM/MeOH (1/1) 100:0 to0:100). The product containing fractions were combined and concentratedunder reduced pressure and further purified by reverse phase preparativeHPLC (RP-HPLC acidic 1: 5 to 95% B in 20 min with a plateau at 95% for 1min). The product containing fractions were combined, basified with aqsat NaHCO₃, extracted twice with DCM, dried through a phase separatorand concentrated under reduced pressure to give the title compound.

LC-MS: Rt=1.06 min; MS m/z [M+H]⁺ 290.3; LC-MS 1

Step 5: 3-(benzyloxy)-4-fluoro-6-methylpicolinic acid

Ethyl 3-(benzyloxy)-4-fluoro-6-methylpicolinate (2.05 g, 5.95 mmol) wasdissolved in 1,4-dioxane (5 mL) and NaOH 1M in water (12.0 mL, 12.0mmol) was added and was stirred at 50° C. for 45 minutes. The RM wasconcentrated under reduced pressure and the residue was mixed with water(30 mL), aq 1M HCl (25 mL) and DCM (30 mL). The aqueous layer was washedwith DCM (3×20 mL). The combined organic layers were dried through aphase separator and concentrated under reduced pressure to give thetitle compound.

LC-MS: Rt=0.56 min; MS m/z [M+H]⁺ 262.2, m/z [M−H]⁻ 260.1; UPLC-MS 1

Intermediate CY: 5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid Step1: 5-(benzyloxy)-4,6-dichloropyrimidine

To a stirred solution of 4,6-dichloropyrimidin-5-ol (29.5 g, 179 mmol)in DMF (100 mL) was added K₂CO₃ (32.1 g, 232 mmol) then(bromomethyl)benzene (23.4 mL, 197 mmol) at RT and the RM was stirred atRT for 3 hours. The RM was diluted with EtOAc/water, extracted once withEtOAc and the organic layer was washed with brine, dried over Na₂SO₄ andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (RediSep Column: Silica 330 g, eluentheptane:EtOAc 100:0 to 87:13). The product containing fractions werecombined and concentrated under reduced pressure to afford the titlecompound as a white solid.

LC-MS: Rt=1.18 min; no mass observed; UPLC-MS 1

Step 2: 5-(benzyloxy)-4-chloro-6-methylpyrimidine

To a stirred solution of 5-(benzyloxy)-4,6-dichloropyrimidine (32.5 g,127 mmol), K₃PO₄ (81.0 g, 382 mmol) and Pd(dppf)Cl₂.DCM (5.20 g, 6.36mmol) in toluene (350 mL) and water (100 mL) was added methyl boronicacid (9.17 g, 153 mmol) in 1,4-dioxane (45 mL) at 105° C. and thereaction was stirred at 105° C. for 18 hours. Methyl boronic acid (9.17g, 153 mmol) was added again and the RM was stirred at 105° C. for 8hours. Methyl boronic acid (9.17 g, 153 mmol) was added again and the RMwas stirred at 105° C. for 4 hours. The RM was diluted with EtOAc/water,extracted twice with EtOAc and the combined organic layers were washedwith water and brine, dried over Na₂SO₄ and concentrated under reducedpressure. The crude product was purified by column chromatography(RediSep Column: Silica 330 g, eluent heptane:EtOAc 100:0 to 77:23). Theproduct containing fractions were combined and concentrated underreduced pressure to give the title compound as a yellow oil.

LC-MS: Rt=1.03 min; MS m/z [M+H]⁺ 235.2/237.2; UPLC-MS 1

Step 3: methyl 5-(benzyloxy)-6-methylpyrimidine-4-carboxylate

A solution of 5-(benzyloxy)-4-chloro-6-methylpyrimidine (25.9 g, 110mmol), Pd(dppf)Cl₂.DCM (4.51 g, 5.52 mmol) and Et₃N (30.8 mL, 221 mmol)in MeOH (25 mL) was stirred at 50° C. under 10 bar of CO for 40 hours.The RM was filtered through celite and concentrated under reducedpressure. The residue was triturated with DCM and the solid was filteredoff. The filtrate was purified by column chromatography (eluentheptane:EtOAc 100:0 to 60:40). The product containing fractions werecombined and concentrated under reduced pressure to give the titlecompound as a white solid.

LC-MS: Rt=0.81 min; MS m/z [M+H]⁺ 259.1; UPLC-MS 1

Step 4: 5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid

To a stirred solution of methyl5-(benzyloxy)-6-methylpyrimidine-4-carboxylate (22.9 g, 88.0 mmol) inTHF (100 mL) and MeOH (100 mL) was added NaOH 2N in water (100 mL, 200mmol) at RT and the RM was stirred at RT for 5 minutes. THF and MeOHwere removed under reduced pressure, then the resulting aqueous residuewas acidified to pH 3 with 2N HCl and the mixture was filtered to givethe title compound as a white solid.

LC-MS: Rt=0.43 min; MS m/z [M+H]⁺ 245.2, m/z [M−H]⁻ 243.1; UPLC-MS 1

Intermediate CZ: 3-methoxy-4-(trifluoromethoxy)picolinic acid Step 1:4-(bromodifluoromethoxy)-2-chloro-3-methoxypyridine

The flask was flame-dried in vacuo and backfilled with argon. The dryargon flushed flask was charged with NaH (1.05 g, 39.3 mmol). Thecontent was suspended in anhydrous DMF (45 mL) and2-chloro-3-methoxypyridin-4-ol (3.00 g, 17.9 mmol) was introducedportionwise. Then the RM was heated up to 60° C. for 1 hour, cooled toRT and left to stir for another 1 hour. The RM was cooled to 0° C. and asolution of dibromodifluoromethane (6.01 mL, 62.5 mmol) in anhydrous DMF(20 mL) was added dropwise with 0.8 mL/min. Then the bath was removedand the resulting RM was stirred at RT for 1.75 hours. The reaction washeated to 60° C. and stirred at 60° C. for 14.25 hours. The reaction waspartitioned between water (190 mL) and EtOAc (70 mL). The organic layerwas collected and the aqueous layer was back-extracted with EtOAc (3×70mL). The organic layers were combined, washed with brine (210 mL) anddried through a phase separator. The mixture was concentrated underreduced pressure. The residue was adsorbed onto Isolute and purified bycolumn chromatography (Buchi® FlashPure ID HP silica cartridge 220 g,eluent heptane:EtOAc 100:0 to 85:15). The product containing fractionswere combined and concentrated under reduced pressure to give the titlecompound as a colourless oil.

LC-MS: Rt=1.10 min; MS m/z [M+H]⁺ 288.0/290.0/292.0; UPLC-MS 1

Step 2: 2-chloro-3-methoxy-4-(trifluoromethoxy)pyridine

The flask was flame-dried in vacuo and backfilled with argon. The dryargon flushed flask was charged with4-(bromodifluoromethoxy)-2-chloro-3-methoxypyridine (890 mg, 3.02 mmol)and anhydrous DCM (30 mL). To the colorless solution was addedportionwise silver tetrafluoroborate (2.48 g, 12.1 mmol). The RM wasstirred at RT for 17.5 hours. The RM was filtered over a pad of celite.To the filtrates were added water (100 mL). The organic layer wascollected and the aqueous layer was back-extracted with DCM (3×30 mL).The organic layers were combined, washed with brine (120 mL) and driedthrough a phase separator. The filtrate was concentrated under reducedpressure. The residue was adsorbed onto Isolute and purified by columnchromatography (Buchi® FlashPure ID HP silica cartridge 25 g, eluentheptane:EtOAc 100:0 to 85:15). The product containing fractions werecombined and concentrated under reduced pressure to give the titlecompound as a colourless oil.

LC-MS: Rt=1.02 min; MS m/z [M+H]⁺ 228.1/230.1; UPLC-MS 1

Step 3: ethyl 3-methoxy-4-(trifluoromethoxy)picolinate

The reactor was charged with2-chloro-3-methoxy-4-(trifluoromethoxy)pyridine (440 mg, 1.90 mmol),Pd(dppf)Cl₂.DCM (77.0 mg, 95.0 μmol), Et₃N (800 μL, 5.68 mmol) andanhydrous EtOH (10 mL). The autoclave was subjected to three cycles ofevacuation-backfilling with argon. Subsequently it was filled with 15bar of CO at RT and then heated up to 80° C. for 15 hours. ISOLUTE®Si-TMT (1.93 g, 947 μmol) was introduced and the suspension was stirredfor 1 hour at 40° C. The RM was filtered over a pad of Celite. Thefiltrate was concentrated under reduced pressure. The residue wasadsorbed onto Isolute and purified by column chromatography (Buchi®FlashPure ID HP silica cartridge 40 g, eluent heptane:EtOAc 100:0 to75:25). The product containing fractions were combined and concentratedunder reduced pressure to give the title compound as a colourless oil.

LC-MS: Rt=0.90 min; MS m/z [M+H]⁺ 266.1; UPLC-MS 1

Step 4: 3-methoxy-4-(trifluoromethoxy)picolinic acid

Ethyl 3-methoxy-4-(trifluoromethoxy)picolinate (355 mg, 1.31 mmol) wasdissolved in 1,4-dioxane (8.75 mL) and the clear colorless solution wastreated with NaOH 1M in water (1.44 mL, 1.44 mmol). The resulting RM wasstirred at RT for 14.75 hours. The reaction was partitioned betweenwater (15 mL) and Et₂O (25 mL). The organic layer was discarded. Theaqueous layer was collected and frozen in a mixture of acetone/dryice.After freeze drying under high pressure the title compound was affordedas a sodium salt.

LC-MS: Rt=0.43 min; MS m/z [M+H]⁺ 238.1, m/z [M−H]⁻ 236.1; UPLC-MS 1

Intermediate DA: 3-(benzyloxy)-4-fluoro-5-methylpicolinic acid Step 1:2-bromo-5-methyl-4-nitropyridin-3-ol

2-Bromo-5-methylpyridin-3-ol (1.50 g, 7.58 mmol) was suspended insulfuric acid (8.42 mL). The resulting dark brown suspension was cooledat 0° C. and then HNO₃ (66%) (729 μL, 10.6 mmol) was introduced. Theresulting RM was left to react at 0° C. for 30.75 hours. HNO₃ (66%) (104mL, 1.52 mmol) was added, then it was stirred for 45 hours. The RM waspoured into ice. The resulting mixture was partitioned between water (50mL) and EtOAc (100 mL). The organic layer was collected and the aqueouslayer was back-extracted with EtOAc (3×40 mL). The organic layers werecombined, washed with brine (50 mL) and dried through a phase separator.The filtrate was concentrated under reduced pressure. The residue wasadsorbed onto Isolute and purified by column chromatography (Redisep® HPsilica cartridge 120 g, eluent heptane:EtOAc 100:0 to 55:45). Theproduct containing fractions were combined and concentrated underreduced pressure to give the title compound as a yellow solid.

LC-MS: Rt=0.46 min; MS m/z [M−H]⁻ 231.0/233.0; UPLC-MS 1

Step 2: 3-(benzyloxy)-2-bromo-5-methyl-4-nitropyridine

The flask was flame-dried in vacuo and backfilled with argon. The dryargon flushed flask was charged with2-bromo-5-methyl-4-nitropyridin-3-ol (422 mg, 1.78 mmol) and K₂CO₃ (743mg, 5.32 mmol). The contents were suspended in anhydrous DMF (5.92 mL)and the resulting RM was treated with (bromomethyl)benzene (280 μL, 2.31mmol). The RM was stirred at RT for 22.5 hours. The reaction waspartitioned between water (40 mL) and EtOAc (30 mL). The organic layerwas collected and the aqueous layer was back-extracted with EtOAc (3×20mL). The organic layers were combined, washed with brine (40 mL) anddried through a phase separator. The filtrate was concentrated underreduced pressure. The residue was adsorbed onto Isolute and purified bycolumn chromatography (Buchi® FlashPure ID HP silica cartridge 40 g,eluent heptane:EtOAc 100:0 to 90:10). The product containing fractionswere combined and concentrated under reduced pressure to give the titlecompound as a white solid.

LC-MS: Rt=1.26 min; MS m/z [M+H]⁺ 323.2/325.2, m/z [M−H]⁻ 321.1/323.1;UPLC-MS 1

Step 3: 3-(benzyloxy)-2-bromo-4-fluoro-5-methylpyridine

The flask was flame-dried in vacuo and backfilled with argon. The dryargon flushed vial was charged with3-(benzyloxy)-2-bromo-5-methyl-4-nitropyridine (539 mg, 1.64 mmol) andanhydrous DMF (5.45 mL). To the clear stirred solution was added TBAF 1Min THF (2.45 mL, 2.45 mmol) and the resulting RM was stirred at RT for7.5 hours. TBAF 1M in THF (490 μL, 490 μmol) was introduced. The RM wasstirred at RT for 14.75 hours. The reaction was partitioned betweenwater (30 mL) and EtOAc (25 mL). The organic layer was collected and theaqueous layer was back-extracted with EtOAc (3×20 mL). The organiclayers were combined, washed with brine (40 mL) and dried through aphase separator. The filtrate was concentrated under reduced pressure.The residue was adsorbed onto Isolute and purified by columnchromatography (Buchi® FlashPure ID HP silica cartridge 40 g, eluentheptane:DCM/MeOH (8/2) 100:0 to 90:10). The product containing fractionswere combined and concentrated under reduced pressure to give the titlecompound as a colourless oil.

LC-MS: Rt=1.27 min; MS m/z [M+H]⁺ 296.2/298.1, m/z [M−H]⁻ 294.0/295.9;UPLC-MS 1

Step 4: ethyl 3-(benzyloxy)-4-fluoro-5-methylpicolinate

The reactor was charged with3-(benzyloxy)-2-bromo-4-fluoro-5-methylpyridine (160 mg, 351 μmol),Pd(dppf)Cl₂.DCM (14.3 mg, 18.0 μmol), Et₃N (148 μL, 1.05 mmol) andanhydrous EtOH (10 mL). The autoclave was subjected to three cycles ofevacuation-backfilling with argon. Subsequently it was filled with 15bar of CO at RT and then heated at 80° C. for 18 hours. The RM wastreated with ISOLUTE® Si-TMT (358 mg, 176 μmol). The suspension wasstirred for 1 hour at 40° C. and filtered over a pad of celite. Themixture was concentrated under reduced pressure. The residue wasadsorbed onto Isolute and purified by column chromatography (Buchi®FlashPure ID HP silica cartridge 12 g, eluent heptane:EtOAc 100:0 to75:25). The product containing fractions were combined and concentratedunder reduced pressure to give the title compound as a colourless oil.

LC-MS: Rt=1.06 min; MS m/z [M+H]⁺ 290.2; UPLC-MS 1

Rebenzylation:

A dry argon flushed flask was charged with ethyl3-(benzyloxy)-4-fluoro-5-methylpicolinate (38.0 mg, 131 μmol) and K₂CO₃(22.0 mg, 158 μmol). The contents were suspended in anhydrous DMF (438μL) and the resulting mixture was treated with (bromomethyl)benzene(17.5 μL, 144 μmol). The RM was stirred at RT for 1.75 hours. Thereaction was partitioned between water (10 mL) and EtOAc (5 mL). Theorganic layer was collected and the aqueous layer was back-extractedthree times with EtOAc. The organic layers were combined, washed withbrine (25 mL) and dried through a phase separator. The filtrate wasconcentrated under reduced pressure and dried under HV to give the titlecompound as a colorless oil.

LC-MS: Rt=1.06 min; MS m/z [M+H]⁺ 290.2; UPLC-MS 1

Step 5: 3-(benzyloxy)-4-fluoro-5-methylpicolinic acid

To a colorless solution of ethyl3-(benzyloxy)-4-fluoro-5-methylpicolinate (40.0 mg, 131 μmol) dissolvedin 1,4-dioxane (876 μL) was added NaOH 1M in water (144 μL, 144 μmol).The resulting light yellow solution was stirred at RT for 2.75 hours.The light yellow solution was diluted with water and extracted with Et₂O(25 mL). The organic layer was discarded. The aqueous layer was frozenand lyophilized to give the title compound as a white solid.

LC-MS: Rt=0.68 min; MS m/z [M+H]⁺ 262.2, m/z [M−H]⁻ 260.2; UPLC-MS 1

Intermediate DB: 5-hydroxy-6-methylpyrimidine-4-carboxylic acid Step 1:4-chloro-5-methoxy-6-methylpyrimidine

4,6-Dichloro-5-methoxypyrimidine (50.0 g, 279 mmol) was dissolved in THF(400 mL) under nitrogen and cooled to 5° C. Methylmagnesium chloride 3M(102 mL, 307 mmol) was added dropwise. The RM was stirred at 5° C. for 1hour, then it was quenched by addition of HCl 1N (250 mL) and extractedtwice with TBME (2×250 mL). The combined organic layers were washed withbrine (250 mL) and concentrated under reduced pressure. The residue wastreated with MeOH to give the title compound in solution.

¹H NMR (400 MHz, DMSO-d₆) δ 8.66 (s, 1H), 3.86 (s, 3H), 2.50 (s, 3H)

Step 2: methyl 5-methoxy-6-methylpyrimidine-4-carboxylate

To a solution of 4-chloro-5-methoxy-6-methylpyrimidine (70.0 g, 441mmol) in MeOH (350 mL) were added Pd(dppf)Cl₂.DCM (21.6 g, 26.5 mmol)and Et₃N (122 mL, 883 mmol). Then the mixture was stirred at 60° C. for16 hours under CO atmosphere (0.3-0.5 MPa). After cooling to RT thesolids were collected by filtration and washed with MeOH. After thefiltrate was concentrated under reduced pressure, IPAc (350 mL) andwater (350 mL) were added. The organic phase and aqueous phase wereseparated, then the aqueous phase was extracted by IPAc (350 mL). Afterconcentration, the residue was purified by column chromatography (silicaglas column, eluent heptane:EtOAc 10:1, isocratic). The productcontaining fractions were combined and concentrated under reducedpressure to give the title compound as an oil.

¹H NMR (400 MHz, DMSO-d₆) δ 8.85 (s, 1H), 3.93 (s, 3H), 3.84 (s, 3H),1.17 (s 3H).

Step 3: 5-hydroxy-6-methylpyrimidine-4-carboxylic acid

Methyl 5-methoxy-6-methylpyrimidine-4-carboxylate (22.0 g, 116 mmol) wasdissolved in HBr (≥40% in water) (94.0 mL, 696 mmol). The reaction washeated and stirred at 40° C. for 10 hours. The solution was treated withHI (≥57% in water) (92.0 mL, 696 mmol) and stirred for further 6 hours.The pH was adjusted to 3-4 with NaOH (50% in water) at 0-20° C. Themixture was filtered to receive a yellow solid. The crude product wastaken into water (150 mL) and HCl (37% in water) (44.0 mL, 536 mmol) wasadded. The suspension was heated at 60° C. for 2 hours, then it wasfiltered. The obtained solid was dried under vacuum to give the titlecompound as a white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 11.71 (s, 2H), 8.66 (s, 1H), 2.47 (s, 3H).

Intermediate DC: ethyl 2-(piperazin-1-yl)acetate Step 1: tert-butyl4-(2-ethoxy-2-oxoethyl)piperazine-1-carboxylate

Tert-butyl piperazine-1-carboxylate (1.00 g, 5.37 mmol) was taken in ACN(10 mL) and ethyl 2-bromoacetate (1.35 g, 8.05 mmol) and K₂CO₃ (2.23 g,16.1 mmol) were added and the RM was stirred at RT for 16 hours. The RMwas diluted with water and extracted with EtOAc. The crude product waspurified by column chromatography (Silica gel column: Silica 12 g,eluent hexane:EtOAc 95:5 to 90:10). The product containing fractionswere combined and concentrated under reduced pressure to give the titlecompound.

LC-MS: Rt=1.19 min; MS m/z [M+H]⁺ 273.3; UPLC-MS 13

Step 2: ethyl 2-(piperazin-1-yl)acetate

Tert-butyl 4-(2-ethoxy-2-oxoethyl)piperazine-1-carboxylate (500 mg, 1.45mmol) was taken in 1,4-dioxane (1 mL) and HCl 4N in 1,4-dioxane (10.0mL, 40.0 mmol) was added and the RM was stirred at RT for 1 hour. The RMwas concentrated under reduced pressure and washed with Et₂O to give thetitle compound.

LC-MS: Rt=0.15 min; MS m/z [M+H]⁺ 173.1; UPLC-MS 12

Intermediate DD: 5-fluoro-2-methyl-4-(trifluoromethyl)aniline Step 1:5-fluoro-2-iodo-4-(trifluoromethyl)aniline

3-Fluoro-4-(trifluoromethyl)aniline (2.00 g, 11.2 mmol) was dissolved inDMF (10 mL) and cooled to 0° C. Then NIS (3.52 g, 15.6 mmol) was addedand the mixture was stirred at 0° C. for 1.25 hours. Water, aq satNaHCO₃ and DCM were added. The aqueous layer was washed twice with DCM.The combined organic layers were dried through a phase separator andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (RediSep Column: Silica 80 g, eluent DCM:DCM/MeOH(9/1) 100:0 to 0:100). The product containing fractions were combined,concentrated under vacuum and dried under HV. The resulting product wasfurther purified in 3 portions by reverse phase preparative HPLC(RP-HPLC acidic 1: 20 to 80% B in 20 min and 2×RP-HPLC acidic 1: 25 to85% B in 20 min). The product containing fractions were combined,basified with aq sat NaHCO₃, extracted twice with DCM, dried through aphase separator and concentrated under reduced pressure to give thetitle compound.

LC-MS: Rt=1.14 min; MS m/z [M−H]⁻ 303.9; UPLC-MS 4

Step 2: 5-fluoro-2-methyl-4-(trifluoromethyl)aniline

5-Fluoro-2-iodo-4-(trifluoromethyl)aniline (1.67 g, 5.48 mmol) wasdissolved in 1,4-dioxane (10 mL) and2,4,6-trimethyl-1,3,5,2,4,6-trioxatriborinane (2.50 mL, 17.9 mmol),Pd(PPh₃)₄ (290 mg, 251 μmol) and K₂CO₃ (3.50 g, 25.3 mmol) were added.The mixture was degassed and flushed with argon and stirred at 80° C.for 4 hours. The RM was filtered, the filter cake was washed with1,4-dioxane and the filtrate was concentrated under reduced pressure.Water, aq sat NaHCO₃ and DCM were added to the filtrate and the aqueouslayer was washed twice with DCM. The combined organic layers were driedthrough a phase separator and concentrated under reduced pressure. Inaddition water, aq sat NaHCO₃ and DCM were added to the previous filtercake. The aqueous layer was washed twice with DCM, the combined organiclayers were dried through a phase separator and concentrated underreduced pressure. The crude product obtained from the filtrate waspurified by reverse phase preparative HPLC (RP-HPLC acidic 1: 10 to 90%B in 25 min). The crude product obtained from the filter cake waspurified in 5 portions by reverse phase preparative HPLC (5×RP-HPLCacidic 1: 30 to 90% B in 25 min). The product containing fractions wereall combined, basified with aq sat NaHCO₃, extracted twice with DCM,dried through a phase separator and concentrated under reduced pressureto give the title compound.

LC-MS: Rt=1.06 min; no mass observed; UPLC-MS 4

Intermediate DE: 2-chloro-5-fluoro-6-(trifluoromethyl)pyridin-3-amine

2-Chloro-5-fluoropyridin-3-amine (1.00 g, 6.82 mmol) andbis(((trifluoromethyl)sulfinyl)oxy)zinc (4.52 g, 13.7 mmol) were mixedin DCM (11 mL) and water (3.8 mL). Then 2-hydroperoxy-2-methylpropane(2.83 mL, 20.5 mmol) was added and the RM was stirred at RT overnight.Water and 5% aq NaHCO₃ was added. The mixture was extracted 3 times withDCM, the organic phase was filtered through Celite, dried through aphase separator and concentrated under reduced pressure. The crudeproduct was purified by column chromatography (RediSep Column: Silica 40g, eluent cyclohexane:EtOAc 100:0 to 50:50). The product containingfractions were combined and concentrated under reduced pressure to givethe title compound.

LC-MS: Rt=0.88 min; MS m/z [M+H]⁺ 215.1/217.1, m/z [M−H]⁻ 213.1/215.1;UPLC-MS 1

Intermediate DF: 2-methyl-4-(pentafluorosulfanyl)aniline Step 1:2-bromo-4-(pentafluorosulfanyl)aniline

4-(Pentafluorosulfanyl)aniline (135 mg, 614 μmol) was dissolved in DCM(4 mL) and cooled to 0° C. Then1,3-dibromo-5,5-dimethylimidazolidine-2,4-dione (90.3 mg, 316 μmol) wasadded. The mixture was stirred at 0° C. for 15 minutes. Water (10 mL),aq sat NaHCO₃ (10 mL) and DCM (10 mL) were added. The aqueous layer waswashed twice with DCM (2×10 mL). The combined organic layers were driedthrough a phase separator and concentrated under reduced pressure togive the title compound.

LC-MS: Rt=1.19 min; no mass observed; UPLC-MS 1

Step 2: 2-methyl-4-(pentafluorosulfanyl)aniline

2-Bromo-4-(pentafluorosulfanyl)aniline (1.40 g, 4.37 mmol) was mixedwith Pd(PPh₃)₄ (257 mg, 222 μmol), K₂CO₃ (2.72 g, 19.7 mmol) andsuspended in 1,4-dioxane (40 mL) Then2,4,6-trimethyl-1,3,5,2,4,6-trioxatriborinane (1.83 mL, 13.1 mmol) wasadded and the mixture was stirred at 80° C. for 5.5 hours, then at RTovernight. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL) wereadded. The aqueous layer was washed twice with DCM (2×10 mL). Thecombined organic layers were dried through a phase separator andconcentrated under reduced pressure. The crude product was purified byreverse phase preparative ISCO (RediSep Column: C18 130 g, eluentwater+0.1% TFA:ACN 100:0 to 0:100). The product containing fractionswere combined, concentrated under reduced pressure and lyophilizedovernight to give the title compound as a yellow oil.

LC-MS: Rt=1.05 min; no mass observed; UPLC-MS 1

Intermediate DG: 5-chloro-2-methyl-6-(trifluoromethyl)pyridin-3-amine

5-Chloro-2-methylpyridin-3-amine (500 mg, 3.44 mmol) andbis(((trifluoromethyl)sulfinyl)oxy)zinc (2.73 g, 8.25 mmol) were mixedin CHCl₃ (10 mL) and water (3.33 mL). 2-Hydroperoxy-2-methylpropane 70%in water (1.43 mL, 10.3 mmol) was added and the RM was stirred at 55° C.for 23 hours. The RM was cooled to RT and diluted with water (10 mL).The organic layer was washed with water (10 mL), dried through a phaseseparator and concentrated under reduced pressure. The crude product wasadsorbed onto Isolute and purified by column chromatography (RediSepColumn: Silica 80 g, eluent DCM:MeOH 100:0 to 85:15). The productcontaining fractions were combined and concentrated under reducedpressure to give the title compound.

LC-MS: Rt=0.95 min; MS m/z [M+H]⁺ 211.1/213.1, m/z [M−H]⁻ 209.0/211.1;UPLC-MS 3

Intermediate DH:rac-2-(4-methoxycyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolaneStep 1: rac-4-methoxycyclohex-1-en-1-yl trifluoromethanesulfonate

To the stirred solution of rac-4-methoxycyclohexan-1-one (5.00 g, 39.0mmol) in dry THF (50 mL) was added LDA (39.0 mL, 78.0 mmol) dropwise at−78° C. The RM was stirred at −78° C. for 30 minutes and then1,1,1-trifluoro-N-phenyl-N-((trifluoromethyl)sulfonyl)methanesulfonamide(20.9 g, 58.5 mmol) in dry THF (50 mL) was added. The RM was stirred at−78° C. for 30 minutes, then at RT for 3 hours. The RM was quenched withaq sat NH₄Cl and extracted three times with EtOAc. The combined organiclayers were washed with water, dried over Na₂SO₄, filtered andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (Silica gel column: Silica 24 g, eluenthexane:EtOAc 100:0 to 99:1). The product containing fractions werecombined, concentrated under reduced pressure and dried under HV to givethe title compound.

Step 2:rac-2-(4-methoxycyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

To the stirred solution of rac-4-methoxycyclohex-1-en-1-yltrifluoromethanesulfonate (4.50 g, 17.3 mmol) in 1,4-dioxane (30 mL)were added 4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane)(6.59 g, 25.9 mmol) and KOAc (5.09 g, 51.9 mmol). The RM was degassedwith argon for 30 minutes. Pd(dppf)Cl₂.DCM (706 mg, 865 μmol) was addedand the RM was degassed with argon for 15 minutes. The RM was stirred at90° C. for 16 hours. The RM was poured onto ice water and extractedtwice with EtOAc. The organic layer was dried over Na₂SO₄ andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (Silica gel column: Silica 24 g, eluenthexane:EtOAc 100:0 to 99:1). The product containing fractions werecombined, concentrated under reduced pressure and dried under HV to givethe title compound as a colorless oil.

Intermediate DI: tert-butyl6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydropyridine-1(2H)-carboxylateStep 1: tert-butyl6-(((trifluoromethyl)sulfonyl)oxy)-3,4-dihydropyridine-1(2H)-carboxylate

To NaHMDS 1M in THE (47.4 mL, 47.4 mmol) in THE (400 mL) at −78° C.under nitrogen, was added tert-butyl 2-oxopiperidine-1-carboxylate (9.44g, 47.4 mmol) in THE (50 mL) over 10 minutes. The resulting mixture wasstirred at −78° C. for 45 minutes. Then1,1,1-trifluoro-N-phenyl-N-((trifluoromethyl)sulfonyl)methanesulfonamide(16.9 g, 47.4 mmol) in THE (40 mL) was added over 10 minutes and stirredfor 1 hour. The RM was allowed to warm slowly to RT overnight, then itwas stirred at RT for 24 hours. The RM was partitioned between TBME (400mL) and NaOH 0.1M (200 mL). The organic layer was separated, washed withbrine (100 mL), dried over Na₂SO₄, filtered and evaporated in vacuo. Thecrude product was purified by column chromatography (RediSep Column:Silica 220 g, eluent heptane:EtOAc:NH₃ 90:10:1 to 85:15:1). The productcontaining fractions were combined and concentrated under reducedpressure to give the title compound.

¹H NMR (400 MHz, DMSO-d₆) δ 5.54 (t, J=3.8 Hz, 1H), 3.49 (m, 2H), 2.24(m, 2H), 1.67 (m, 2H), 1.44 (s, 9H)

Step 2: tert-butyl6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydropyridine-1(2H)-carboxylate

To BISPIN (9.79 g, 38.5 mmol), PdCl₂(PPh₃)₂ (966 mg, 1.38 mmol),triphenylphosphane (722 mg, 2.75 mmol) and K₂CO₃ (7.13 g, 51.6 mmol) wasadded tert-butyl6-(((trifluoromethyl)sulfonyl)oxy)-3,4-dihydropyridine-1(2H)-carboxylate(11.4 g, 34.4 mmol) in 1,4-dioxane (150 mL). The RM was degassed,flushed with argon and heated at 90° C. for 3 hours then at 70° C.overnight. After cooling to RT the mixture was diluted with TBME (200mL) and washed with water (100 mL). The organic phase was dried overNa₂SO₄, filtered and concentrated under reduced pressure. The crudeproduct was purified by column chromatography (eluent heptane:EtOAc5:1). The product containing fractions were combined and concentratedunder reduced pressure to give 2 batches of the title compound as palebrown oils.

LC-MS: Rt=1.20 min; no mass observed, UPLC-MS: 1

Intermediate DJ: tert-butyl3-fluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydropyridine-1(2H)-carboxylateStep 1: tert-butyl 5-fluoro-2-oxopiperidine-1-carboxylate

5-Fluoropiperidin-2-one (513 mg, 4.38 mmol), di-tert-butyl dicarbonate(2.03 mL, 8.76 mmol) and DMAP (53.5 mg, 438 μmol) were stirred in ACN(25 mL) and THF (10 mL) at RT for 3 days. The RM was concentrated underreduced pressure. The crude product was adsorbed onto Isolute andpurified by column chromatography (RediSep Column: Silica 12 g, eluentheptane:EtOAc 100:0 to 40:60). Fractions containing TLC R_(f) 0.44(heptane:EtOAc, 1:1, KMnO₄ visualization) were combined and evaporatedin vacuo to give the title compound as a pale yellow oil.

¹H NMR (600 MHz, DMSO-d₆) δ 5.14 (m, 1H), 3.98 (m, 1H), 3.67 (m, 1H),2.41 (m, 2H), 2.17 (m, 1H), 1.96 (m, 1H), 1.44 (s, 9H)

Step 2: tert-butyl3-fluoro-6-(((trifluoromethyl)sulfonyl)oxy)-3,4-dihydropyridine-1(2H)-carboxylate

To KHMDS 0.5M in toluene (2.39 mL, 1.20 mmol) in THF (10 mL) at −78° C.under nitrogen was added dropwise a solution of tert-butyl5-fluoro-2-oxopiperidine-1-carboxylate (208 mg, 957 μmol) in THF (2 mL).The RM was allowed to stir at −78° C. for 1.75 hours. Then addeddropwise bis(trifluoromethanesulfonyl)aniline (428 mg, 1.20 mmol) in THF(2 mL) and stirred at −78° C. for 10 minutes. The RM was allowed to warmto RT and stirred for 1 hour at RT. NaOH 1M (15 mL) was added and themixture was extracted with Et₂O (2×30 mL). The organic layers werecombined, washed with brine (10 mL), dried over MgSO₄ and K₂CO₃,filtered and evaporated in vacuo to give a yellow oil (540 mg). Thecrude product was purified by column chromatography (RediSep Column:Silica 12 g, eluent heptane:EtOAc:NH₃ 95:5:0.1 to 0:100:1). The productcontaining fractions were combined and concentrated under reducedpressure to give the title compound as a yellow oil.

LC-MS: Rt=1.20 min; MS m/z [M−H]⁻ 348.0; UPLC-MS 1

Step 3: tert-butyl3-fluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydropyridine-1(2H)-carboxylate

To BISPIN (83.0 mg, 326 μmol), PdCl₂(PPh₃)₂ (7.64 mg, 10.9 μmol),triphenylphosphane (5.71 mg, 22.0 μmol) and K₂CO₃ (56.4 mg, 408 μmol)was added tert-butyl3-fluoro-6-(((trifluoromethyl)sulfonyl)oxy)-3,4-dihydropyridine-1(2H)-carboxylate(95.0 mg, 272 μmol) in 1,4-dioxane (1.5 mL). The RM was flushed withargon and heated at 70° C. for 18 hours. The RM was partitioned betweenDCM (8 mL) and water (3 mL). The organic layer was dried through a phaseseparator and concentrated under reduced pressure to give the titlecompound as a dark brown oil.

Intermediate DK: 3,6-dihydro-2H-pyran-4-carbaldehyde

According to ref. Org. Lett. 2014, 16, 4142-4145.

To a mixture of tetrahydro-4H-pyran-4-one (30.0 g, 300 mmol) and water(300 mL), was added NaCN (15.4 g, 315 mmol) at 5° C. followed by NaHSO₄until a pH=4-5 was reached. The reaction was stirred at 10° C. for 1hour, then NaCl (17.5 g, 300 mmol) was added at 25° C. followed by2-MeTHF. The organic layers were separated and the aqueous layer wasextracted twice with 2-MeTHF. The combined organic layers were washedwith brine, dried over Na₂SO₄, concentrated under reduced pressure andswitched the solvent to toluene (300 mL) to give4-hydroxytetrahydro-2H-pyran-4-carbonitrile. Pyridine (48.5 mL, 599mmol) was added at 65° C., followed by a slow addition of POCl3 (27.9mL, 300 mmol). The RM was stirred at 65° C. for 1 hour, then it wascooled to RT and water was added. The layers were separated, and theaqueous layer was extracted twice with toluene. The combined organiclayers were washed with brine, dried over Na₂SO₄ and concentrated underreduced pressure to give 3,6-dihydro-2H-pyran-4-carbonitrile. Theresidue was mixed with toluene (300 mL) and DIBAL-H (46.9 g, 330 mmol)was added at −10° C. The reaction was stirred at −10° C. for 1 hour,then HCl 4M was added. The two layers were separated, and the aqueouslayer was extracted twice with DCM. The combined organic layers werewashed with brine and dried over Na₂SO₄. The organic solution was thenconcentrated under reduced pressure to give the title compound assolution in toluene (not stable when concentrated).

¹H NMR (400 MHz, DMSO-d₆) δ 9.46 (s, 1H), 7.05 (m, 1H), 4.33 (m, 2H),3.69 (m, 2H), 2.16 (m, 2H).

Intermediate DK: 3,6-dihydro-2H-pyran-4-carbaldehyde Step 1: methyltetrahydro-2H-pyran-4-carboxylate

According to ref. WO2013/66729, 2013, A1

Into a solution of oxane-4-carboxylic acid (50.0 mg, 380 μmol) in MeOH(10 mL) was added thionyl chloride (46.0 mg, 390 μmol) dropwise withstirring at RT. The resulting solution was stirred at RT for 3 hours.The RM was concentrated under reduced pressure to give the titlecompound, which was used directly for next step without furtherpurification (brown oil).

¹H NMR (400 MHz, DMSO-d₆) δ 3.81 (m, 2H), 3.62 (s, 3H), 3.34 (m, 2H),2.59 (m, 1H), 1.74 (m, 2H), 1.56 (m, 2H).

Step 2: methyl 4-iodotetrahydro-2H-pyran-4-carboxylate

To a solution of DIPEA (528 mL, 3.03 mol) in THE (3.5 L) was addeddropwise n-BuLi 2M in hexane (1.52 L, 3.03 mmol) at −10° C.˜−5° C. TheRM was stirred at −10° C.˜−5° C. for 30 minutes. Methyltetrahydro-2H-pyran-4-carboxylate (370 g, 2.43 mol) in THE (700 mL) wasadded dropwise into the RM at −70° C. The RM was stirred for at −70° C.for 1 hour. I₂ (1.23 g, 4.85 mol) in THF (2.4 L) was added dropwise intothe RM at −70° C. The RM was stirred for at −70° C. for 1 hour. Themixture was added dropwise into HCl 1N (1.5 L) and TBME (1.5 L) and itwas stirred for 30 minutes. The organic was washed with aq 30% Na₂O₃S₂(1.8 L) and brine (700 mL). The organic layer was concentrated underreduced pressure to give the title compound.

Step 3: methyl 3,6-dihydro-2H-pyran-4-carboxylate

To a solution of methyl 4-iodotetrahydro-2H-pyran-4-carboxylate (500 g,1.61 mol) in TBME (10 L) was added dropwise DBU (486 mL, 3.22 mol) atRT. The RM was stirred for at RT 2 hours, then it was filtered and tothe filtrate was added HCl 9% (1.7 L). The mixture was stirred for 30minutes. The pH of the organic was adjusted to 7 by washing with brine(2×1.5 L). The combined aqueous phase was extracted with TBME (1.5 L).After concentration, the residue was purified by distillation collects55° C. of distillate to give the title compound.

¹H NMR (400 MHz, DMSO-d₆) δ 6.92 (m, 1H), 4.21 (m, 2H), 3.69 (m, 5H),2.25 (m, 2H).

Step 4: (3,6-dihydro-2H-pyran-4-yl)methanol

Methyl 3,6-dihydro-2H-pyran-4-carboxylate (50.0 g, 288 mmol) was mixedin THE (900 mL) under nitrogen. The solution was cooled to −10° C.,followed by addition of DIBAL-H 1M in hexane (721 mL, 721 mmol)dropwise. The RM was stirred at 0° C. for 2 hours, then it was quenchedby addition of 3N Rochelle's salt solution (700 mL) and MeOH (117 mL).The biphasic solution was stirred for 16 hours, and the organic phasewas isolated. The aqueous phase was extracted with DCM:MeOH 10:1 (5×200mL) and the combined organic phases were concentrated under reducedpressure to give the title compound as a liquid.

¹H NMR (400 MHz, DMSO-d₆) δ 5.61 (m, 1H), 4.74 (m, 1H), 4.02 (m, 2H),3.82 (m, 2H), 3.67 (m, 2H), 1.97 (m, 2H).

Step 5: 3,6-dihydro-2H-pyran-4-carbaldehyde

To a solution of DMP (122 g, 288 mol) in IPAc (600 mL) under nitrogenwas added dropwise (3,6-dihydro-2H-pyran-4-yl)methanol (36.0 g, 262mmol) at 0° C. The RM was stirred at RT for 3 minutes. The suspensionwas filtered and washed with IPAc (36 mL). The pH of the filtrate wasadjusted to 8 by washing with 10% Na₂CO₃ (500 mL). The aqueous phase wasextracted with IPAc (2×150 mL). The combined organic phase wasconcentrated under reduced pressure to ½ volume to give the titlecompound as a solution in IPAc.

¹H NMR (400 MHz, DMSO-d₆) δ 9.46 (s, 1H), 7.05 (m, 1H), 4.33 (m, 2H),3.69 (m, 2H), 2.16 (m, 2H).

Intermediate DL: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-iodoacetamideStep 1: 2-chloro-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

2-Chloro-4-(trifluoromethyl)aniline (18.5 g, 95.0 mmol) was dissolved inDCM (180 mL) at 0° C. A solution of 2-chloroacetyl chloride (10.7 g,95.0 mmol) in DCM (40 mL) was added dropwise over 15 minutes. After 30minutes at 0° C. the RM was warmed to RT. The white suspension wasstirred at RT overnight. The suspension was filtered and washed withDCM. The filtrate was concentrated under reduced pressure and driedunder HV to give the title compound as a white solid.

LC-MS: Rt=1.14 min; MS m/z [M−H]⁻ 270.1/272.1/274.0; UPLC-MS 1

Step 2: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-iodoacetamide

2-Chloro-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (15.8 g, 58.2mmol) was dissolved in acetone (215 mL), KI (10.6 g, 64.0 mmol) wasadded and the RM was stirred at reflux for 2.25 hours. The RM was cooledto RT and the suspension was filtered. The cake was washed with acetoneand DCM. The filtrate was concentrated under reduced pressure and driedunder HV to give the title compound.

LC-MS: Rt=1.13 min; MS m/z [M−H]⁻ 362.0/364.0; UPLC-MS 1

Intermediate DM:2-bromo-N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)acetamide Step 1:5-chloro-2-methyl-4-(trifluoromethyl)aniline

2-Bromo-5-chloro-4-(trifluoromethyl)aniline (2.70 g, 9.84 mol) wassuspended in 1,4-dioxane (30 mL) and methylboronic acid (883 mg, 14.8mol) and K₂CO₃ (3.40 g, 24.6 mmol) were added. The RM was degassed withargon for 15 minutes and Pd(dppf)Cl₂.DCM (402 mg, 492 μmol) was addedand the RM was stirred at 80° C. for 16 hours. The RM was filteredthrough Celite and washed with 1,4-dioxane. The filtrate wasconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (Silica gel column: Silica 12 g, eluenthexane:EtOAc 100:0 to 90:10). The product containing fractions werecombined and concentrated under reduced pressure to give the titlecompound.

LC-MS: Rt=1.56 min; MS m/z [M+H]⁺ 210.1/212.1; UPLC-MS 11

Step 2: 2-bromo-N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

5-Chloro-2-methyl-4-(trifluoromethyl)aniline (1.50 g, 6.94 mmol) wassuspended in DCM (20 mL) was mixed at 0° C. with 2-bromoacetyl bromide(665 μL, 7.64 mmol) and DMAP (848 mg, 6.94 mmol) and the RM was stirredat RT for 12 hours. The RM was diluted with DCM and washed with water.The organic layer was dried over Na₂SO₄ and concentrated under reducedpressure, washed with Et₂O and dried under HV to give the titlecompound.

LC-MS: Rt=1.58 min; MS m/z [M+H]⁺ 330.0/332.0/334.0; UPLC-MS 11

Intermediate DN:2-bromo-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide Step 1:2-bromo-5-fluoro-4-(trifluoromethyl)aniline

To a stirred solution of1-bromo-4-fluoro-2-nitro-5-(trifluoromethyl)benzene (5.00 g, 17.4 mmol)in EtOH (50 mL) and AcOH (50 mL) was added Fe dust (2.91 g, 52.1 mmol)and the RM was stirred at 80° C. for 2 hours. The solvents wereevaporated and NaOH 5N was added. The mixture was extracted with EtOAc,the organic layer was dried and concentrated under reduced pressure togive the title compound.

LC-MS: Rt=1.58 min; no mass observed; UPLC-MS 11

Step 2: 5-fluoro-2-methyl-4-(trifluoromethyl)aniline

2-Bromo-5-fluoro-4-(trifluoromethyl)aniline (1.90 g, 7.22 mmol) wasdissolved in 1,4-dioxane (50 mL) and degassed using argon. Thenmethylboronic acid (1.08 g, 18.0 mmol), K₂CO₃ (2.99 g, 21.7 mmol) andPd(dppf)Cl₂.DCM (589 mg, 722 μmol) were added and the RM was stirred at80° C. for 12 hours. The RM was filtered through Celite. The filtratewas concentrated under reduced pressure. The crude product was purifiedby column chromatography (Silica gel column: Silica 24 g, eluenthexane:EtOAc 100:0 to 95:5). The product containing fractions werecombined, concentrated under reduced pressure and dried under HV to givethe title compound.

LC-MS: Rt=1.55 min; no mass observed; UPLC-MS 11

Step 3: 2-bromo-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

5-Fluoro-2-methyl-4-(trifluoromethyl)aniline (670 mg, 3.33 mmol) wassuspended in DCM (10 mL) and DMAP (407 mg, 3.33 mmol) was added at 0° C.The RM was stirred at 0° C. for 10 minutes, then 2-bromoacetyl bromide(348 μL, 4.00 mmol) was added. The RM was stirred at RT for 12 hours.Water was added to the RM. The organic layer was dried over Na₂SO₄ andconcentrated under reduced pressure to give the title compound.

LC-MS: Rt=1.45 min; MS m/z [M+H]⁺ 314.0/316.0; UPLC-MS 13

Intermediate DO:2-bromo-N-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)acetamide

2-Chloro-5-fluoro-4-(trifluoromethyl)aniline (400 mg, 1.87 mmol) wassuspended in DCM (5 mL). 2-Bromoacetyl bromide (179 μL, 2.06 mmol) andDMAP (229 mg, 1.87 mmol) were added at 0° C. and the RM was stirred atRT for 12 hours. The RM was cooled to 0° C., water was added and thesuspension was filtered. The cake was washed with hexane and dried underreduced pressure. The crude product was purified by columnchromatography (Silica gel column: Silica 12 g, eluent DCM:MeOH 100:0 to98:2). The product containing fractions were combined and concentratedunder reduced pressure to give the title compound.

LC-MS: Rt=2.28 min; MS m/z [M+H]⁺ 333.9/335.9/337.9; UPLC-MS 13

Intermediate DP: 2-bromo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To 2-methyl-4-(trifluoromethyl)aniline (5.00 g, 28.5 mmol) in DCM (100mL) was added dropwise 2-bromoacetyl bromide (3.73 mL, 42.8 mmol) at 0°C., followed by DIPEA (15.0 mL, 86.0 mmol) and the resulting blacksolution was stirred at RT for 30 minutes. 2-Bromoacetyl bromide (1.87mL, 21.4 mmol) was added to the reaction and it was stirred for 1 hour.The RM was diluted with DCM (100 mL) and water (100 mL). The biphasicmixture was given in a separating funnel and shaken for 1 minute. Theorganic layer was dried over Na₂SO₄ and concentrated to give a blacksolution. The black solution was purified by column chromatography(eluent heptane:EtOAc 100:0 to 50:50). The product containing fractionswere combined and concentrated under reduced pressure to give the titlecompound as a brown solid.

LC-MS: Rt=1.04 min; MS m/z [M−H]⁻ 293.9/295.9; UPLC-MS 4

Intermediate DQ:2-bromo-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide

2-Chloro-6-(trifluoromethyl)pyridin-3-amine (10.0 g, 50.9 mmol) wassuspended in DCM (150 mL) and DMAP (6.22 g, 50.9 mmol) was added at 0°C. The RM was stirred at 0° C. for 10 minutes, then 2-bromoacetylbromide (5.32 mL, 61.1 mmol) was added and the RM was stirred at RT for12 hours. The RM was diluted with DCM, washed with water and the organiclayer was dried over Na₂SO₄ and concentrated under reduced pressure. Thecrude product was purified by column chromatography (Silica gel column:Silica 40 g, eluent hexane:EtOAc 100:0 to 90:10). The product containingfractions were combined, concentrated under reduced pressure and driedunder HV to give the title compound.

LC-MS: Rt=1.52 min; MS m/z [M+H]⁺ 316.9/318.9/320.9; UPLC-MS 12

Intermediate DR: 2-bromo-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide

To the stirred solution of 2-fluoro-4-(trifluoromethyl)aniline (4.00 g,22.3 mmol) in DCM (20 mL) were added 2-bromoacetyl bromide (2.14 mL,24.6 mmol) and DMAP (2.73 g, 22.3 mmol) at 0° C. The RM was stirred atRT for 16 hours. The RM was concentrated under reduced pressure. Waterwas added and it was extracted with 5% MeOH in DCM. It was washed twicewith HCl 1N and the combined organic layers were dried over Na₂SO₄,filtered and concentrated under reduced pressure to give the titlecompound.

HPLC: Rt=7.705 min; HPLC 2

Intermediate DS:2-bromo-N-(4-chloro-2-methyl-5-(trifluoromethyl)phenyl)acetamide

4-Chloro-2-methyl-5-(trifluoromethyl)aniline (1.00 g, 4.77 mmol) wassuspended in DCM (20 mL), 2-bromoacetyl bromide (457 μL, 5.25 mmol) andDMAP (583 mg, 4.77 mmol) were added at 0° C. and the RM was stirred atRT for 12 hours. The RM was diluted with DCM and washed with water. Theorganic layer was dried over Na₂SO₄, concentrated under reducedpressure, washed with Et₂O and dried to give the title compound as awhite solid.

LC-MS: Rt=1.56 min; MS m/z [M+H]⁺ 329.8/331.9/333.9; UPLC-MS 11

Intermediate DT: 2-bromo-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

2-Chloro-4-(trifluoromethyl)aniline (3.00 g, 15.3 mmol) was suspended inDCM (30 mL) and DMAP (1.87 g, 15.3 mmol) was added at 0° C. and stirredfor 10 minutes. 2-Bromoacetyl bromide (3.72 g, 18.4 mmol) was added andthe RM was stirred at RT for 12 hours. The organic layer was washed withHCl 1N. The layers were separated and the organic layer was dried overNa₂SO₄ and concentrated under reduced pressure to give the titlecompound.

LC-MS: Rt=1.59 min; MS m/z [M+H]⁺ 316.0/318.0/320.0; UPLC-MS 11

Intermediate DU:N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-iodoacetamide Step 1:2-chloro-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide

2-Chloro-6-(trifluoromethyl)pyridin-3-amine (3.70 g, 18.8 mmol) wasdissolved in DCM (20 mL) and 2-chloroacetyl chloride (1.80 mL, 22.6mmol) was added to the yellow solution, followed by Et₃N (6.52 mL, 47.1mmol). The RM turned immediately into a dark solution. The RM wasstirred at RT for 1.5 hours. Water (30 mL) and DCM (30 mL) were added.The aqueous layer was washed twice with DCM (2×30 mL). The combinedorganic layers were dried through a phase separator and concentratedunder reduced pressure. The crude product was purified by columnchromatography (RediSep Column: Silica 120 g, eluent DCM:DCM/MeOH (9/1)100:0 to 90:10). The product containing fractions were combined,concentrated under reduced pressure and dried under HV to afford thetitle compound as an orange solid.

LC-MS: Rt=0.98 min; MS m/z [M−H]⁻ 271.1/273.1/275.1; UPLC-MS 3

Step 2: N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-iodoacetamide

2-Chloro-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide (5.46 g,16.0 mmol) and KI (6.50 g, 39.2 mmol) were suspended in ACN (50 mL) andstirred at 82° C. for 45 minutes. The RM was cooled to RT and filtered.The cake was washed with ACN (3×50 mL). The filtrate was concentratedunder reduced pressure. The crude product was dissolved in a smallamount of DCM and purified by column chromatography (RediSep Column:Silica 120 g, eluent DCM. The product containing fractions werecombined, concentrated under reduced pressure and dried under HV toafford the title compound as a pale beige solid.

LC-MS: Rt=0.98 min; MS m/z [M−H]⁻ 362.9/364.9; UPLC-MS 3

Intermediate DV: 2-iodo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: 2-chloro-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

2-Methyl-4-(trifluoromethyl)aniline (10.0 g, 56.0 mmol) was dissolved inDCM (93 mL) at 0° C. Then 2-chloroacetyl chloride (4.72 mL, 58.7 mmol)was added, followed by Et₃N (17.1 mL, 123 mmol) (violet solution). After30 minutes was the RM warmed to RT and it was stirred at RT for 1.2hours. The RM was extracted with DCM (3×200 mL) and with water (2×60mL). The organic layer was dried through a phase separator andconcentrated under reduced pressure. The crude product was suspended ina small amount of DCM and filtered. The cake was washed with a smallamount of DCM and Et₂O. The cake was dried under reduced pressureovernight. The filtrate was concentrated under reduced pressure. Theresidue was adsorbed onto Isolute and purified by column chromatography(RediSep Column: Silica 80 g, eluent heptane:EtOAc 50:50 to 0:100. Theproduct containing fractions were combined and concentrated underreduced pressure and combined with the cake to give the title compound.

LC-MS: Rt=1.01 min; MS m/z [M−H]⁻ 250.1/252.1; UPLC-MS 4

Step 2: 2-iodo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

2-Chloro-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (11.8 g, 46.8mmol) was dissolved in acetone (173 mL) and KI (10.1 g, 60.9 mmol) wasadded. The RM was stirred at reflux for 2.5 hours. The RM was cooled toRT and the suspension was filtered. The cake was washed with acetone andDCM. The filtrate was concentrated under reduced pressure to give abeige solid. The solid was suspended in DCM and filtered to give thetitle compound as beige solid.

LC-MS: Rt=1.04 min; MS m/z [M−H]⁻ 342.0; UPLC-MS 4

Intermediate DW:2-bromo-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide Step 1:2-methyl-6-(trifluoromethyl)pyridin-3-amine

To the stirred solution of 2-chloro-6-(trifluoromethyl)pyridin-3-amine(5.00 g, 25.4 mmol) in 1,4-dioxane (50 mL) and water (10 mL) were addedmethylboronic acid (2.28 g, 38.2 mmol) and K₂CO₃ (7.03 g, 50.9 mmol) atRT and degassed with nitrogen for 15 minutes. Pd(dppf)Cl₂ (1.04 g, 1.27mmol) was added and the RM was stirred at 100° C. for 12 hours. Water(20 mL) was added and the mixture was extracted with EtOAc (2×20 mL).The organic layer was washed with brine (20 mL), dried over Na₂SO₄ andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (Silica gel column: Silica 12 g, eluenthexane:EtOAc 100:0 to 75:25). The product containing fractions wereconcentrated under reduced pressure and dried under HV to give the titlecompound.

LC-MS: Rt=0.47 min; MS m/z [M+H]⁺ 177.2; UPLC-MS 12

Step 2: 2-bromo-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

To the stirred solution of 2-methyl-6-(trifluoromethyl)pyridin-3-amine(2.40 g, 13.5 mmol) in DCM (30 mL) were added 2-bromoacetyl bromide(2.99 g, 14.8 mmol) and DMAP (1.65 g, 13.5 mmol) and the RM was stirredat RT for 4 hours. Water (20 mL) was added to the RM and it wasextracted with EtOAC (2×20 mL). The organic layer was dried over Na₂SO₄and concentrated under reduced pressure. The crude product was purifiedby column chromatography (Silica gel column: Silica 12 g, eluenthexane:EtOAc 100:0 to 75:25). The product containing fractions wereconcentrated under reduced pressure and dried under HV to give the titlecompound.

LC-MS: Rt=1.29 min; MS m/z [M+H]⁺ 297.0/299.0; UPLC-MS 13

Intermediate DX:2-chloro-N-(4-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide

4-Chloro-6-(trifluoromethyl)pyridin-3-amine (300 mg, 1.53 mmol) wassuspended in ACN (10 mL). K₂CO₃ (211 mg, 1.53 mmol) and 2-chloroacetylchloride (146 μL, 1.83 mmol) were added at 0° C. and the RM was stirredat 70° C. for 12 hours. Water was added and the organic layer was driedover Na₂SO₄ and concentrated under reduced pressure to give the titlecompound.

LC-MS: Rt=1.49 min; MS m/z [M+H]⁺ 273.1/275.1/277.1; UPLC-MS 11

Intermediate DY:2-bromo-N-(4-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

4-Methyl-6-(trifluoromethyl)pyridin-3-amine (210 mg, 1.19 mmol) wassuspended in DCM (3 mL) and DMAP (146 mg, 1.19 mmol) was added at 0° C.and the RM was stirred for 10 minutes. 2-bromoacetyl bromide (125 μL,1.43 mmol) was added and the RM was stirred at RT for 12 hours. Waterwas added. The organic layer was washed with water, dried over Na₂SO₄and concentrated under reduced pressure to give the title compound.

LC-MS: Rt=1.73 min; MS m/z [M+H]⁺ 297.2/299.2; UPLC-MS 13

Intermediate DZ:N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)-2-iodoacetamide Step 1:2-chloro-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

3-Fluoro-2-methyl-4-(trifluoromethyl)aniline (1.19 g, 6.14 mmol) wasdissolved in DCM (25 mL) and 2-chloroacetyl chloride (538 μL, 6.75 mmol)was added at 0° C., followed by Et₃N (1.70 mL, 12.3 mmol) and stirred atRT for 15 minutes. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL)were added. The aqueous layer was washed twice with DCM (2×10 mL). Thecombined organic layers were dried through a phase separator andconcentrated under reduced pressure to give the title compound. Thecrude product still contained ca 34% of starting material.

LC-MS: Rt=1.00 min; MS m/z [M−H]⁻ 268.1/270.1; UPLC-MS 1

LC-MS: Rt=4.98 min; MS m/z [M−H]⁻ 268.1/270.0; UPLC-MS 2

Step 2: N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)-2-iodoacetamide

2-Chloro-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (1.30g, 4.82 mmol) were mixed in acetone (20 mL) and KI (880 mg, 5.30 mmol)was added. The RM was stirred at 60° C. for 30 minutes. The reaction wasstopped and filtered. The cake was washed several times with acetone.The filtrate was concentrated under reduced pressure. As shown in theLCMS in the step before the reaction contains ca 34% aniline. For thatreason the concentrated filtrate was dissolved in DCM (10 mL) and2-chloroacetyl chloride (258 μL, 3.24 mmol) was added, followed by Et₃N(898 μL, 6.48 mmol). The RM was stirred at RT for 20 minutes. LCMSshowed a small amount of aniline left. Water (10 mL), aq sat NaHCO₃ (10mL) and DCM (10 mL) were added. The aqueous layer was washed twice withDCM (2×100 mL). The combined organic layers were dried through a phaseseparator and concentrated under reduced pressure. The crude productfrom step 2 was restarted with KI (550 mg, 3.31 mmol) in acetone (10 mL)at 60° C. for 1 hour. The reaction was stopped and filtered. The cakewas washed several times with acetone. The filtrate was concentratedunder reduced pressure to give the title compound.

LC-MS: Rt=1.05 min; MS m/z [M−H]⁻ 360.1; UPLC-MS 1

Intermediate EA:2-chloro-N-(2-chloro-4-(pentafluorosulfanyl)phenyl)acetamide Step 1:2-chloro-4-(pentafluorosulfanyl)aniline

4-(Pentafluorosulfanyl)aniline (2.00 g, 9.13 mmol) and NCS (1.28 g, 9.58mmol) were diluted in DMF (18 mL) and the resulting RM was then stirredat 80° C. for 1 hour. The reaction was stopped and cooled down. The RMwas adsorbed onto Isolute and purified by column chromatography (RediSepColumn: Silica 80 g, eluent cyclohexane:cyclohexane/TBME (1/1) 100:0 to40:60). The product containing fractions were combined and evaporated todryness to give the title compound as beige, slightly pink crystals.

LC-MS: Rt=0.98 min; no mass observed; UPLC-MS 7

Step 2: 2-chloro-N-(2-chloro-4-(pentafluorosulfanyl)phenyl)acetamide

2-Chloro-4-(pentafluorosulfanyl)aniline (404 mg, 1.59 mmol) wasdissolved in DCM (5 mL) and 2-chloroacetyl chloride (152 μL, 1.91 mmol)was added, followed by Et₃N (551 μL, 3.98 mmol). The RM was stirred atRT for 3 hours. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL)were added. The aqueous layer was washed twice with DCM (2×10 mL). Thecombined organic layers were dried through a phase separator andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (RediSep Column: Silica 24 g, eluent DCM:DCM/MeOH(1/1) 100:0 to 70:30). The product containing fractions were combined,concentrated under reduced pressure and dried under HV to afford thetitle compound as a beige solid.

LC-MS: Rt=1.22 min; MS m/z [M−H]⁻ 328.0/330.0/332.0; UPLC-MS 1

Intermediate EB: 2-bromo-N-(5-fluoro-4-formyl-2-methylphenyl)acetamideStep 1: tert-butyl (5-fluoro-4-formyl-2-methylphenyl)carbamate

To the stirred solution of 4-bromo-2-fluoro-5-methylbenzaldehyde (3.50g, 16.1 mmol) in toluene (70 mL) were added tert-butyl carbamate (3.78g, 32.3 mmol) and Cs₂CO₃ (10.5 g, 32.3 mmol). The RM was degassed withnitrogen for 15 minutes. BINAP (502 mg, 806 μmol) and Pd(OAc)₂ (181 mg,806 μmol) were added and the RM was stirred at 100° C. for 14 hours.Water (50 mL) was added and it was extracted with DCM (2×50 mL). Theorganic layer was washed with brine (20 mL), dried over Na₂SO₄ andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (Silica gel column: Silica 12 g, eluent DCM:MeOH100:0 to 99:1). The product containing fractions were combined andconcentrated under reduced pressure to give the title compound.

LC-MS: Rt=1.7 min; MS m/z [M+H]⁺ 254.1; UPLC-MS 12

Step 2: 4-amino-2-fluoro-5-methylbenzaldehyde

To the stirred solution of tert-butyl(5-fluoro-4-formyl-2-methylphenyl)carbamate (3.20 g, 12.5 mmol) in Et₂O(15 mL) was added HCl 2M in Et₂O (30.0 mL, 60.0 mmol) and the RM wasstirred at RT for 4 hours. The RM was concentrated under reducedpressure and washed with Et₂O to give the title compound.

LC-MS: Rt=0.81 min; MS m/z [M+H]⁺ 154.3; UPLC-MS 13

Step 3: 2-bromo-N-(5-fluoro-4-formyl-2-methylphenyl)acetamide

To the stirred solution of 4-amino-2-fluoro-5-methylbenzaldehyde (1.90g, 11.4 mmol) in DCM (30 mL) at 0° C. were added 2-bromoacetyl bromide(2.30 g, 11.4 mmol) and DMAP (1.39 g, 11.4 mmol) and the RM was stirredat RT for 4 hours. Water (20 mL) was added and the RM was extracted withEtOAc (2×20 mL). The organic layer was washed with brine (20 mL), driedover Na₂SO₄ and concentrated under reduced pressure. The crude productwas purified by column chromatography (Silica gel column: Silica 40 g,eluent hexane:EtOAc 100:0 to 70:30). The product containing fractionswere combined and concentrated to give the title compound.

LC-MS: Rt=1.42 min; MS m/z [M+H]⁺ 274.1/276.1; UPLC-MS 13

Intermediate EC:2-iodo-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide Step 1:2-chloro-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

2-Methyl-6-(trifluoromethyl)pyridin-3-amine (2.00 g, 11.4 mmol) wasdissolved in DCM (19 mL). At 0° C. 2-chloroacetyl chloride (958 μL, 11.9mmol) was added. The solution turned into a white suspension. Et₃N (3.46mL, 25.0 mmol) was added. The white suspension turned into a brownsolution, which was stirred at 0° C. for 25 minutes. Then it was allowedto warm to RT. After 1.2 hours 2-chloroacetyl chloride (300 mg, 2.63mmol) was added and the RM was stirred at RT for 17.5 hours.2-Chloroacetyl chloride (200 mg, 1.75 mmol) was added again and the RMwas continued stirring at RT for 1.5 hours. The RM was extracted withDCM (3×80 mL) and water (2×10 mL). The combined organic layers weredried through a phase separator and concentrated under reduced pressure.The residue was adsorbed onto Isolute and purified by columnchromatography (RediSep Column: Silica 40 g, eluent DCM:MeOH 100:0 to80:20). All product containing fractions were combined to give the titlecompound as a white solid.

LC-MS: Rt=0.83 min; MS m/z [M+H]⁺ 253.1/255.1, m/z [M−H]⁻ 251.1/253.1;UPLC-MS 3

Step 2: 2-iodo-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

2-Chloro-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide (2.28 g,7.94 mmol) was dissolved in acetone (30 mL). KI (1.71 g, 10.3 mmol) wasadded to the pale brown solution and the RM was stirred at reflux for1.3 hours. The RM was cooled to RT and the suspension was filtered. Thecake was washed with acetone (2×20 mL). The filtrate was concentratedunder reduced pressure to give a beige solid, which was suspended in DCM(20 mL) and filtered. The cake was washed with DCM (3×20 mL) and thefiltrate was concentrated under reduced pressure to give the titlecompound as a slight yellow solid.

LC-MS: Rt=0.88 min; MS m/z [M+H]⁺ 345.0, m/z [M−H]⁻ 342.9; UPLC-MS 3

Intermediate ED:N-(5-fluoro-2-methyl-6-(trifluoromethyl)pyridin-3-yl)-2-iodoacetamideStep 1: 5-fluoro-2-methyl-6-(trifluoromethyl)pyridin-3-amine

5-Fluoro-2-methylpyridin-3-amine.HCl (533 mg, 3.28 mmol) andbis(((trifluoromethyl)sulfinyl)oxy)zinc (2.58 g, 7.78 mmol) were mixedin DCM (5 mL) and water (1.7 mL). Then 2-hydroperoxy-2-methylpropane(1.36 mL, 9.83 mmol) was added. The mixture was stirred at RT for 5hours. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL) were added.The aqueous layer was washed twice with DCM (2×10 mL). The combinedorganic layers were dried through a phase separator and concentratedunder reduced pressure. The crude product was purified in 3 portions byreverse phase preparative HPLC (3×RP-HPLC acidic 1: 5 to 100% B in 20min). The product containing fractions were combined, basified with aqsat NaHCO₃, extracted twice with DCM, dried through a phase separatorand concentrated under reduced pressure to give the title compound.

LC-MS: Rt=0.89 min; MS m/z [M+H]⁺ 195.1, m/z [M−H]⁻ 193.0; UPLC-MS 3

Step 2:2-chloro-N-(5-fluoro-2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

5-Fluoro-2-methyl-6-(trifluoromethyl)pyridin-3-amine (436 mg, 2.25 mmol)was dissolved in DCM (3 mL) and 2-chloroacetyl chloride (232 μL, 2.91mmol) was added dropwise, followed by Et₃N (685 μL, 4.94 mmol). The RMwas stirred at RT for 15 minutes. 2-Chloroacetyl chloride (100 μL, 1.26mmol) was added. The RM was stirred at RT for 15 minutes. Water (10 mL),aq sat NaHCO₃ (10 mL) and DCM (10 mL) were added. The aqueous layer waswashed twice with DCM (2×10 mL). The combined organic layers were driedthrough a phase separator and concentrated under reduced pressure togive the title compound.

LC-MS: Rt=0.98 min; MS m/z [M+H]⁺ 271.2/273.1, m/z [M−H]⁻ 269.0/271.0;UPLC-MS 3

Step 3:N-(5-fluoro-2-methyl-6-(trifluoromethyl)pyridin-3-yl)-2-iodoacetamide

2-Chloro-N-(5-fluoro-2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide(771 mg, 2.71 mmol) and KI (899 mg, 5.41 mmol) were suspended in ACN (15mL) and stirred at 82° C. for 30 minutes. Water (10 mL), aq sat NaHCO₃(10 mL) and DCM (10 mL) were added. The aqueous layer was washed twicewith DCM (2×10 mL). The combined organic layers were dried through aphase separator and concentrated under reduced pressure to give thetitle compound.

LC-MS: Rt=1.04 min; MS m/z [M+H]⁺ 363.0, m/z [M−H]⁻ 360.9; UPLC-MS 3

Intermediate EE:2-chloro-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide

3-Fluoro-2-methyl-4-(trifluoromethyl)aniline (295 mg, 1.53 mmol) wasdissolved in DCM (5 mL) and 2-chloroacetyl chloride (122 μL, 1.53 mmol)was added, followed by Et₃N (423 μL, 3.05 mmol). The RM was stirred atRT for 1 hour. 2-Chloroacetyl chloride (122 μL, 1.53 mmol) was added,followed by Et₃N (423 μL, 3.05 mmol). The RM was stirred at RT for 1hour. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL) were added.The aqueous layer was washed twice with DCM (2×10 mL). The combinedorganic layers were dried through a phase separator and concentratedunder reduced pressure to give the title compound.

LC-MS: Rt=1.01 min; MS m/z [M−H]⁻ 268.2/270.1; UPLC-MS 1

Intermediate EF:2-chloro-N-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)acetamide

2-Chloro-5-fluoro-4-(trifluoromethyl)aniline (205 mg, 960 μmol) wasdissolved in DCM (2 mL) and 2-chloroacetyl chloride (80.0 μL, 1.01 mmol)was added, followed by Et₃N (400 μL, 2.89 mmol). The RM turned from acolorless solution to a brown solution. The RM was stirred at RT for 30minutes. 2-Chloroacetyl chloride (120 μL, 1.51 mmol) was added and theRM was stirred at RT for 30 minutes. Water (10 mL), aq sat NaHCO₃ (10mL) and DCM (10 mL) were added. The aqueous layer was washed twice withDCM (2×10 mL). The combined organic layers were dried through a phaseseparator and concentrated under reduced pressure to give the titlecompound.

LC-MS: Rt=1.22 min; MS m/z [M−H]⁻ 288.0/290.0/292.0; UPLC-MS 1

Intermediate EG: (S)-((2-fluoro-4-iodobutoxy)methyl)benzene Step 1:(R)-1-(benzyloxy)pent-4-en-2-ol

To a colorless solution of (R)-2-((benzyloxy)methyl)oxirane (9.29 mL,59.7 mmol) in THF (200 mL) was added CuBr-DMS complex (12.5 g, 59.7mmol). The brown fine suspension was cooled to −45° C. Thenvinylmagnesium bromide 1M in THE (209 mL, 209 mmol) was added dropwisewithin 30 minutes at −45° C. to −35° C. After the addition the darkbrown suspension was stirred at −45° C. for 20 minutes. The RM wasallowed to warm to RT and stirred for 1 hr. The brown RM was warmed to−15° C. and quenched with aq sat NH₄Cl (500 mL) and stirred at 0° C. for30 minutes. The RM was filtered over celite, washed with TBME (5×200 mL)and the blue biphasic filtrate was extracted. The organic phase wasextracted twice with brine (2×300 mL). The aqueous layers were washedtwice with TBME (2×300 mL). The combined organic phases were dried overNa₂SO₄, filtered and concentrated under reduced pressure at 45° C. togive the title compound as a yellow oil.

LC-MS: Rt=0.90 min; no mass observed; UPLC-MS 1

Step 2: (S)-(((2-fluoropent-4-en-1-yl)oxy)methyl)benzene

To a pale yellow solution of (R)-1-(benzyloxy)pent-4-en-2-ol (12.0 g,62.4 mmol) in benzotrifluoride 547948 (180 mL) were added DIPEA (164 mL,936 mmol) and triethylamine trihydrofluoride (61.0 ml, 374 mmol) at RT.A slight exotherm increased the internal temperature to a maximum of 29°C. 1,1,2,2,3,3,4,4,4-Nonafluorobutane-1-sulfonyl fluoride (11.2 mL, 62.4mmol) was added at 25° C. (exothermic: internal temperature was slowlyincreased to a maximum of 36° C.). The clear yellow brown RM was stirredat RT for 1 hour (outside temperature was set to 20° C.).1,1,2,2,3,3,4,4,4-Nonafluorobutane-1-sulfonyl fluoride (11.2 mL, 62.4mmol) was added to the pale brown RM at RT (slightly exothermic). Theclear pale brown RM was stirred at RT for 1 hour and1,1,2,2,3,3,4,4,4-nonafluorobutane-1-sulfonyl fluoride (11.2 mL, 62.4mmol) was added. The clear pale brown RM was stirred at RT for 1 hour.1,1,2,2,3,3,4,4,4-Nonafluorobutane-1-sulfonyl fluoride (11.2 mL, 62.4mmol) was added. The clear pale brown RM was stirred at RT overnight.The RM was poured into ice-cold HCl 2N (600 mL) and stirred for about 10minutes. Then the phases were separated and the organic phase wasextracted successively with ice-cold HCl 2N (500 mL), aq sat NaHCO₃ (400mL) and water (400 mL). The aqueous layer was washed twice with TBME(2×300 mL). The combined organic phases were dried over Na₂SO₄, filteredand the filtrate was evaporated in vacuuo at 50° C. to give crudeproduct as a brown resin (24.5 g). The crude product was purified bycolumn chromatography (Biotage Sfaer Silica D 60 μm cartridge: 350 g,eluent heptane:EtOAc 100:0 to 60:40). The product containing fractionswere combined and concentrated under reduced pressure to give the titlecompound as a yellow oil.

LC-MS: Rt=1.31 min; no mass observed; UPLC-MS 1

Step 3: (S)-4-(benzyloxy)-3-fluorobutan-1-ol

A pale yellow solution of(S)-(((2-fluoropent-4-en-1-yl)oxy)methyl)benzene (11.5 g, 59.2 mmol) wasdissolved in DCM (400 mL) and MeOH (100 mL). The pale yellow solutionwas cooled in a dry ice/acetone-bath to −74° C. internal temperature.Adjust oxygen to purge the lines (5 minutes). Then ozone generator wasswitched on (200 L/hour O₂, and 100% power). (slightly exothermic;internal temperature was slowly increased to a maximum of −68° C.).After about 13 minutes the RM was blue. Performance of the ozonegenerator adjusted to zero. During the flushing of the generator and theapparatus with argon, sodium borohydride (4.03 g, 107 mmol) was addedportionwise within about 3 minutes to the RM (slightly exothermic;internal temperature was slowly increased to a maximum of −69° C.). TheRM decolorized from blue to colorless. After 5 minutes the cooling bathwas removed and the RM was allowed to warm to RT (ca 45 minutes). The RMwas stirred at RT for 1 hour. The RM was poured onto ice cold HCl 0.5N(500 mL) and stirred for 5 minutes. Then the phases were separated. Theorganic phase was extracted once with aq sat NaHCO₃ (500 mL). Theaqueous layer was washed twice with DCM (2×200 mL). The combined organicphases were dried over Na₂SO₄, filtered and the solvent was removed invacuo at 45° C. to give the title compound as a colorless slightlycloudy oil.

LC-MS: Rt=0.67 min; MS m/z [M+H]⁺ 199.1, m/z [M−H]⁻ 197.9; UPLC-MS 1

Step 4: (S)-((2-fluoro-4-iodobutoxy)methyl)benzene

To a colorless solution of (S)-4-(benzyloxy)-3-fluorobutan-1-ol (10.0 g,50.4 mmol) in THF (230 mL) were added triphenylphosphane (19.9 g, 76.0mmol) and 1H-imidazole (6.87 g, 101 mmol). The colorless solution wascooled in an ice/MeOH bath to −10° C. Then I₂ (18.6 g, 73.1 mmol) wasadded (strong exotherm, temperature rose to 5° C.). The brown RM wasstirred at 0° C. for 2 hours. After 30 minutes a fine yellow suspensionwas formed. The RM was poured onto ice cold aq sat NH₄Cl (500 mL) andTBME (500 mL) and stirred for 5 minutes. Then the phases were separated.The organic phase was extracted once with aq sat NaHCO₃ (500 mL) andbrine (500 mL). The aqueous layer was washed twice with TBME (2×300 mL).The combined organic phases were dried over Na₂SO₄, filtered andconcentrated under reduced pressure at 45° C. until thetriphenylphosphane oxide began to crystallize. Then heptane (250 mL) wasadded slowly. The rest of TBME and THE was evaporated. The yellowsuspension was stirred at 10° C. for 30 minutes. The precipitatedcrystals were filtered off and the solid was washed with heptane. Thecake was discarded. The filtrate was concentrated under reduced pressureat 45° C. to give a yellow oil (16.2 g) The crude product was purifiedby column chromatography (Biotage Sfaer Silica D 60 μm cartridge: 350 g,eluent heptane:DCM 100:0 to 50:50). The product containing fractionswere combined. The impure fractions were purified by columnchromatography (Biotage Sfaer Silica D 60 μm; 350 g, eluent heptane:DCM100:0 to 50:50). The product containing fractions were combined with theones from the first purification and concentrated under reduced pressureto give the title compound as a pale yellow oil.

LC-MS: Rt=1.33 min; no mass observed; UPLC-MS 1

Intermediate EH: tert-butyl4-(1-methoxy-1,3-dioxopentan-2-yl)piperazine-1-carboxylate Step 1:methyl 2-chloro-3-oxopentanoate

To a solution of methyl 3-oxopentanoate (10.4 kg, 80.0 mol) in DCM (67L) was added SO₂Cl₂ (14.0 kg, 104 mol) at RT over 2.5 hours. Thereaction was allowed to warm to RT and stirred for 16 hours. The RM wasconcentrated under reduced pressure and the residue was dissolved in DCM(20 L) and washed with water (10 L), brine (10 L), dried over Na₂SO₄ andfiltered. The filtrate was concentrated under reduced pressure to givethe title compound as a light yellow liquid.

¹H NMR (400 MHz, CDCl₃-d) δ 4.65 (s, 1H), 3.68 (s, 3H), 2.59 (m, 2H),0.96 (t, 3H)

Step 2: tert-butyl4-(1-methoxy-1,3-dioxopentan-2-yl)piperazine-1-carboxylate

To a solution of methyl 2-chloro-3-oxopentanoate (12.1 kg, 53.7 mol) indry ACN (53 L) was added Et₃N (22.3 L, 161 mol) over 1.5 hours, followedby dropwise addition of tert-butyl piperazine-1-carboxylate (10.0 kg,53.7 mol) in ACN (50 L) over 2.5 hours. The reaction was stirred at 60°C. for 16 hours. The RM was filtered and washed with EtOAc (10 L). Thefiltrate was then concentrated under reduced pressure and the residuewas dissolved in EtOAc (45 L) and washed with water (45 L), dried overNa₂SO₄ and filtered. The filtrate was concentrated under reducedpressure and the residue was purified by column chromatography (Silicacolumn 150 mm×800 mm×70 mm, eluent heptane:EtOAc 100:0 to 90:10) to givethe title compound.

HPLC: Rt=3.681/5.513 min; HPLC 4

Intermediate EI: ethyl 2-bromo-3-oxopentanoate

Ethyl 3-oxopentanoate (10.0 g, 65.9 mmol) was dissolved in DCM (100 mL),NBS (12.3 g, 69.2 mmol) was added, followed by TsOH.H₂O (2.53 g, 13.2mmol). The RM was stirred at RT for 2.5 hours. The RM was filteredthrough Hyflo and washed with DCM. The filtrate was extracted with water(3×50 mL). The organic layer was washed with DCM (3×200 mL). Thecombined organic layers were dried through a phase separator andconcentrated under reduced pressure to give the title compound as acolourless liquid.

LC-MS: Rt=0.90 min; MS m/z [M−H]⁻ 221.1/223.1; UPLC-MS 8

Intermediate EJ: tert-butyl4-(1-ethoxy-1,3-dioxopentan-2-yl)piperazine-1-carboxylate

Ethyl 2-bromo-3-oxopentanoate (Intermediate EI) (2.87 g, 7.97 mmol),tert-butyl piperazine-1-carboxylate (8.20 g, 44.0 mmol) and K₂CO₃ (6.61g, 47.8 mmol) were mixed in ACN (50 mL). The suspension was stirred atRT for 30 minutes. The RM was filtered. The cake was washed with ACN.The filtrate was concentrated under reduced pressure. The crude productwas purified by column chromatography (RediSep Column: Silica 80 g,eluent DCM:DCM/MeOH (9/1) 100:0 to 0:100). The product containingfractions were combined, concentrated under reduced pressure and driedunder HV to give the title compound.

LC-MS: Rt=1.11/1.38 min; MS m/z [M+H]⁺ 329.3, m/z [M−H]⁻ 327.2; UPLC-MS8

Intermediate EK: tert-butyl(2R)-4-(1-ethoxy-1,3-dioxopentan-2-yl)-2-methylpiperazine-1-carboxylate

Ethyl 2-bromo-3-oxopentanoate (Intermediate EI) (15.6 g, 62.9 mmol) andtert-butyl (R)-2-methylpiperazine-1-carboxylate (13.2 g, 66.1 mmol) weremixed in ACN (250 mL) and K₂CO₃ (17.4 g, 126 mmol) was added. Thereaction was stirred at RT for 4.5 hours. The suspension was filteredand the cake was washed with ACN. The filtrate was concentrated underreduced pressure. The yellow oil was extracted with DCM (4×200 mL) andwater (2×150 mL). The organic layer was dried through a phase separatorand concentrated under reduced pressure. The residue was adsorbed ontoIsolute and purified by column chromatography (RediSep Column: Silica330 g, eluent heptane:EtOAc 100:0 to 75:25) The product containingfractions were combined and concentrated under reduced pressure to getthe title compound as a colourless oil.

LC-MS: Rt=1.20/1.42 min; MS m/z [M+H]⁺ 343.3, m/z [M−H]⁻ 341.1; UPLC-MS8

Intermediate EL: tert-butyl(3S)-4-(1-ethoxy-1,3-dioxopentan-2-yl)-3-methylpiperazine-1-carboxylate

Ethyl 2-bromo-3-oxopentanoate (Intermediate EI) (5.00 g, 22.4 mmol) andtert-butyl (S)-3-methylpiperazine-1-carboxylate (4.71 g, 23.5 mmol) weremixed in ACN (100 mL) and K₂CO₃ (6.20 g, 44.8 mmol) was added. The RMwas stirred at RT for 3.25 hours. Then it was filtered through Hyflo,washed with ACN and concentrated under reduced pressure. The orange oilwas extracted with DCM (4×100 mL) and water (3×50 mL). The organic layerwas dried through a phase separator and concentrated under reducedpressure. The crude product was adsorbed onto Isolute and purified bycolumn chromatography (RediSep Column: Silica 120 g, eluentheptane:EtOAc 100:0 to 0:100). The product containing fractions werecombined and concentrated under reduced pressure to give the titlecompound as a pale yellow liquid.

LC-MS: Rt=1.14/1.45 min; MS m/z [M+H]⁺ 343.3, UPLC-MS 1

Intermediate EM: tert-butyl(3R)-4-(1-ethoxy-1,3-dioxopentan-2-yl)-3-methylpiperazine-1-carboxylate

Ethyl 2-bromo-3-oxopentanoate (Intermediate EI) (5.00 g, 22.4 mmol) andtert-butyl (R)-3-methylpiperazine-1-carboxylate (4.71 g, 23.5 mmol) weremixed in ACN (100 mL). K₂CO₃ (6.20 g, 44.8 mmol) was added and the RMwas stirred at RT for 3.3 hours. The suspension was filtered throughHyflo and washed with ACN. The filtrate was concentrated under reducedpressure. The residue was extracted with DCM (4×100 mL) and water (3×50mL). The organic layer was dried through a phase separator andconcentrated under reduced pressure. The crude product was adsorbed ontoIsolute and purified by column chromatography (RediSep Column: Silica120 g, eluent heptane:EtOAc 100:0 to 0:100). The product containingfractions were combined and concentrated under reduced pressure to givethe title compound as a pale yellow liquid.

LC-MS: Rt=1.14/1.45 min; MS m/z [M+H]⁺ 343.4, UPLC-MS 1

Intermediate EN: tert-butyl(2R,5S)-4-(1-ethoxy-1,3-dioxopentan-2-yl)-2,5-dimethylpiperazine-1-carboxylate

A beige suspension of tert-butyl(2R,5S)-2,5-dimethylpiperazine-1-carboxylate (24.0 g, 106 mmol), ethyl2-bromo-3-oxopentanoate (Intermediate EI) (25.0 g, 106 mmol) and K₂CO₃(22.1 g, 160 mmol) in ACN (266 mL) was stirred at RT for 8.25 hours. TheRM was filtered. The cake was washed with ACN and discarded. Thefiltrate was diluted with DCM and washed once with water. The aqueouslayer was extracted twice with DCM. The combined organic phases weredried through a phase separator, concentrated under reduced pressure anddried under vacuum (40° C.) to give a brown oil. The crude product wasadsorbed onto Isolute and purified in 2 portions by columnchromatography (RediSep Column: Silica 330 g, eluent heptane:EtOAc 100:0to 30:70) and (RediSep Column: Silica 330 g, eluent heptane:EtOAc 100:0to 20:80). The product containing fractions were combined andconcentrated under reduced pressure to give the title compound.

LC-MS: Rt=1.20/1.49 min; MS m/z [M+H]⁺ 357.1, m/z [M+H]⁺ 355.4; UPLC-MS1

Intermediate EO: tert-butyl(2R)-4-(1-ethoxy-1,3-dioxobutan-2-yl)-2-methylpiperazine-1-carboxylate

To a colorless solution of tert-butyl(R)-2-methylpiperazine-1-carboxylate (10.0 g, 50.0 mmol) in toluene (100mL) was added ethyl 2-chloro-3-oxobutanoate (8.73 mL, 60.0 mmol). Theresulting yellow solution was stirred at 100° C. for 3.25 hours. The RMwas concentrated under reduced pressure and dried under vacuum (40° C.)to give a brown residue. The crude product was adsorbed onto Isolute andpurified by column chromatography (RediSep Column: Silica 220 g, eluentheptane:EtOAc 100:0 to 10:90). The product containing fractions werecombined and concentrated under reduced pressure to give the titlecompound as a yellow oil.

LC-MS: Rt=1.12/1.35 min; MS m/z [M+H]⁺ 329.2, m/z [M+H]⁺ 327.1; UPLC-MS8

Intermediate EP: tert-butyl4-(1-ethoxy-1,3-dioxobutan-2-yl)piperazine-1-carboxylate

To a stirred solution of tert-butyl piperazine-1-carboxylate (150 g, 805mmoL) in ACN (1.5 L) at RT was added K₂CO₃ (223 g, 1.61 mol) and the RMwas stirred for 15 minutes. Then ethyl 2-chloro-3-oxobutanoate (112 mL,809 mmol) was added slowly at the same temperature. The resulting RM wasstirred at RT for 16 hours. The RM was filtered through celite pad. Thecelite pad was washed with EtOAc (2 L). The combined organic layers wereconcentrated under reduced pressure to get crude residue. The residuewas dissolved in EtOAc (3 L) and then washed with ice cold water, brine,dried over Na₂SO₄ and concentrated under reduced pressure to get crudeproduct as pale brown liquid. The crude product was purified by columnchromatography (silica gel, 60-120 mesh, eluent petroleum ether:EtOAc100:0 to 85:15). The pure product containing fractions were combined andconcentrated under reduced pressure to give the title compound as aliquid. The impure fractions were combined and concentrated underreduced pressure. Then they were purified again by column chromatography(silica gel, 60-120 mesh, eluent petroleum ether:EtOAc 100:0 to 85:15).The pure product containing fractions were combined and concentratedunder reduced pressure to give the title compound as a liquid. Bothliquids were mixed, dissolved in DCM and concentrated under reducedpressure to get the title compound as a brown liquid. The liquid wasagain dissolved in DCM, concentrated under reduced pressure. The processwas repeated three times and then dried under vacuum to give the titlecompound as a brown liquid.

HPLC: Rt=11.763 min; HPLC 6

Intermediate EQ: tert-butyl4-(1-methoxy-1,3-dioxopentan-2-yl)-2,3-dimethylpiperazine-1-carboxylate

To a stirred solution of tert-butyl 2,3-dimethylpiperazine-1-carboxylate(26.0 g, 121 mmol) in ACN (250 mL) were added K₂CO₃ (25.2 g, 182 mmol)and ethyl 2-bromo-3-oxopentanoate (Intermediate EI) (27.1 g, 121 mmol)at RT and the RM was stirred at RT for 16 hours. The RM was diluted withEtOAc/water, extracted once with EtOAc and the organic extract waswashed with water and brine, dried over Na₂SO₄ and concentrated underreduced pressure to give the title compound as a yellow oil.

LC-MS: Rt=1.21/1.51 min; MS m/z [M+H]⁺ 357.6; UPLC-MS 1

Intermediate ER: 3-bromo-1H-1,2,4-triazol-5-amine Step 1:3,5-dibromo-1-(methoxymethyl)-1H-1,2,4-triazole

To a solution of NaH (846 g, 21.2 mol, 60%) in DMF (12 L) was added3,5-dibromo-1H-1,2,4-triazole (4.00 kg, 17.6 mol) at 10° C. Theresulting solution was stirred at 10° C. for 1 hour. This was followedby the addition of chloro(methoxy)methane (1.70 kg, 21.2 mol) dropwiseat 20° C. The mixture was stirred at RT overnight. The reaction wasquenched with H₂O (20 L). The resulting solution was extracted with TBME(2×7 L). The combined organic layers were washed with 10% NaCl (2×7 L),dried over Na₂SO₄ and concentrated under reduced pressure at 40° C. Theresidue was triturated with heptane (3 L) to give the title compound asa white solid.

HPLC: Rt=3.042 min; HPLC 4

Step 2: 3-bromo-1-(methoxymethyl)-1H-1,2,4-triazol-5-amine

3,5-Dibromo-1-(methoxymethyl)-1H-1,2,4-triazole (800 g, 2.78 mol) wasdissolved in 25% NH₃.H₂O (2.89 L, 16.3 mol) and MeOH (80 mL) at RT. Themixture was stirred at 120° C. for 18 hours. The mixture was cooled to5˜10° C., the solids were collected by filtration and washed with water(200 mL). The cake was dried under vacuum at 60° C. to give the titlecompound as a white solid.

HPLC: Rt=1.701 min; HPLC 4

Step 3: 3-bromo-1H-1,2,4-triazol-5-amine

To a solution of 3-bromo-1-(methoxymethyl)-1H-1,2,4-triazol-5-amine (329g, 1.45 mol) in MeOH (1.5 L) was added HBr (4.39 kg, 21.7 mol) at RT.The mixture was stirred at 100° C. for 18 hours. The mixture wasadjusted to pH=7.0˜7.5 with 10% NaOH at 20-30° C. and extracted withEtOAc (10×3 L). The combined organic phase was dried over Na₂SO₄ andconcentrated under reduced pressure at 50° C. to give the title compoundas a white solid.

HPLC: Rt=0.702 min; HPLC 4

HPLC: Rt=1.902 min; HPLC 5

Intermediate ER: 3-bromo-1H-1,2,4-triazol-5-amine

To a solution of 1H-1,2,4-triazole-3,5-diamine (300 g, 3.03 mol) inHBr/H₂O (2.4 L) was added dropwise NaNO₂ (313 g, 4.54 mol) in water (782mL) at 0° C. over 1.5 hours. The reaction was allowed to warm to RT andstirred for 1 hour. The reaction was stirred at 100° C. and for 16hours. The RM was cooled to RT, filtered and the pH of the mixture (66batches combined) was adjusted to 4 by addition of 10% NaOH. The mixturewas extracted with EtOAc (2×55 L), dried over Na₂SO₄ and filtered. Theorganic phase was concentrated under reduced pressure to give the titlecompound. The pH of the aqueous phase was adjusted to 7-7.5 by 10% NaOH.Then it was extracted with EtOAc (10×35 L), dried over Na₂SO₄ andfiltered. The organic phase was concentrated under reduced pressure togive the title compound.

HPLC: Rt=1.933 min; HPLC 5

Intermediate ES:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamideStep 1: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-4-(2-((2-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

2-(6-(4-(Tert-butoxycarbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)aceticacid (Intermediate BW—step 3) (120 mg, 253 μmol) and2-methyl-6-(trifluoromethyl)pyridin-3-amine (46.8 mg, 266 μmol) weremixed in EtOAc (1.7 mL). Et3N (175 μL, 1.26 mmol) and T₃P 50% in DMF(301 μL, 506 μmol) were added. The RM was stirred at RT for 30 minutes.Water (5 mL) was added, and the RM was extracted with EtOAc (4×20 mL),water (2×5 mL) and brine (5 mL). The organic layer was dried through aphase separator and concentrated under reduced pressure to give thetitle compound as a beige solid (172 mg, 93% pure, yield: 100%).

LC-MS: Rt=1.01 min; MS m/z [M−H]⁻ 250.1/252.1; UPLC-MS 4

Step 2:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-4-(2-((2-methyl-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(172 mg, 253 μmol) was mixed with DCM (1.7 mL). TFA (390 μL, 5.06 mmol)was added and the RM was stirred at RT for 1 hour. The RM wasconcentrated under reduced pressure. DCM and toluene was added and themixture was concentrated again. This was performed twice. The residuewas dried under HV to give the title compound as a beige solid (187 mg,72% pure, yield: 100%).

LC-MS: Rt=0.72 min; MS m/z [M+H]⁺ 533.3, m/z [M−H]⁻ 531.3; UPLC-MS 4

Intermediate ET:2-(2-bromo-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

To a stirred solution of 5-hydroxy-6-methylpyrimidine-4-carboxylic acid(Intermediate DB) (102 mg, 664 μmol) in DCM (10 mL) was added1-chloro-N,N,2-trimethylprop-1-en-1-amine (113 μL, 857 μmol) and the RMwas stirred at RT for 30 minutes. Then2-(2-bromo-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide(Intermediate BZ—step 2) (300 mg, 553 μmol) was added and the RM wasstirred at RT for 30 minutes. DIPEA (290 μL, 1.66 mmol) in DCM (2 mL)was added dropwise and the RM was stirred at RT for 1 hour. The RM wasdiluted with DCM and water, extracted twice with DCM and the combinedorganic layers were washed with water, dried over Na₂SO₄ andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography (RediSep Column: Silica 24 g, eluent DCM:DCM/MeOH(8/2) 100:0 to 50:50). The product containing fractions were combinedand concentrated to give the title compound (100 mg, 77% pure, yield:21%).

LC-MS: Rt=1.06 min; MS m/z [M+H]⁺ 678.4/680.4; UPLC-MS 1

Intermediate EU: rac-tert-butyl5-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylate

Chiral separation of rac-tert-butyl5-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylate(Intermediate CM-Step 5) (88.0 mg):

Preparative chiral HPLC (instrument: SEPIATEC SFC100; column: OVEN3Chiralpak IB-N 250×30 mm 5 μm; eluent: A: 25% [MeOH+0.05% NH₃] B: 75%scCO₂; flow rate: 80.0 mL/min; detection: 240 nm; injection volume: 1.5mL; Gradient: isocratic A: 25%, B: 75% scCO₂)

Intermediate EV: tert-butyl(1R,6S)-5-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylateor tert-butyl (1S,6R)-5-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylate

First eluting stereoisomer, white solid (40.0 mg, 100% pure, yield: 45%)

Chiral HPLC (C-HPLC 13): Rt=1.44 min, 99.5% ee

LC-MS: Rt=1.37 min; MS m/z [M+H−Boc]⁺ 592.5/594.5, m/z [M−H]⁻690.5/692.4; UPLC-MS 1

LC-MS: Rt=6.80 min; MS m/z [M+H−Boc]⁺ 592.3/594.3, m/z [M−H]⁻690.4/692.4; UPLC-MS 2

Intermediate EW: tert-butyl(1R,6S)-5-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylateor tert-butyl (1S,6R)-5-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylate

Second eluting stereoisomer, white solid (40.0 mg, 100% pure, yield:45%)

Chiral HPLC (C-HPLC 13): Rt=2.05 min, 99.5% ee

LC-MS: Rt=1.37 min; MS m/z [M+H−Boc]⁺ 592.5/594.5, m/z [M−H]⁻690.4/692.4; UPLC-MS 1

LC-MS: Rt=6.77 min; MS m/z [M+H−Boc]⁺ 592.3/594.3, m/z [M−H]⁻690.4/692.3; UPLC-MS 2

Intermediate EX:2-(6-((1S,6R)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamideor2-(6-((1R,6S)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

Tert-butyl(1S,6R)-5-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylateor tert-butyl(1R,6S)-5-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylate(Intermediate EV) (first eluting stereoisomer—40.0 mg, 58.0 μmol) wasmixed with HCl 4N in 1,4-dioxane (1.00 mL, 4.00 mmol) and the RM wasstirred at RT for 1 hour. The RM was diluted with 10% MeOH in DCM andNaHCO₃, extracted twice with DCM and the combined organic layers weredried over Na₂SO₄ and concentrated under reduced pressure to give thetitle compound as a white foam (38.0 mg, 90% pure, yield: 100%).

LC-MS: Rt=0.80 min; MS m/z [M+H]⁺ 592.5/594.5, m/z [M−H]⁻ 590.3/592.4;UPLC-MS 1

Intermediate EY:2-(6-((1S,6R)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamideor2-(6-((1R,6S)-2,5-diazabicyclo[4.2.0]octan-2-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

Tert-butyl(1S,6R)-5-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylateor tert-butyl(1R,6S)-5-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylate(Intermediate EW) (second eluting stereoisomer—40.0 mg, 58.0 μmol) wasmixed with HCl 4N in 1,4-dioxane (1.00 mL, 4.00 mmol) and the RM wasstirred at RT for 1 hour. The RM was diluted with 10% MeOH in DCM andNaHCO₃, extracted twice with DCM and the combined organic layers weredried over Na₂SO₄ and concentrated under reduced pressure to give thetitle compound as a white foam (37.0 mg, 90% pure, yield: 97%).

LC-MS: Rt=0.79 min; MS m/z [M+H]⁺ 592.3/594.3, m/z [M−H]⁻ 590.3/592.3;UPLC-MS 1

Intermediate EZ:N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl-2,2,3,3,5,5,6,6-d8)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

2-(6-Bromo-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide(Intermediate CM—step 4) (500 mg, 892 μmol) andpiperazine-2,2,3,3,5,5,6,6-d8 (403 mg, 4.28 mmol) were mixed in DMSO(300 μL). The RM was stirred at 120° C. in a capped vial for 1.75 hours.The RM was diluted with water (3 mL) and the suspension was stirred atRT for 1 hour. The beige solid was filtered off and redissolved in DCM(20 mL). Then it was filtered through a phase separator and concentratedunder reduced pressure to ca 4 mL. The crude product was purified bycolumn chromatography (RediSep Column: Silica 24 g, eluent DCM:MeOH100:0 to 80:20). The product containing fractions were combined,concentrated, and dried under HV to give the title compound as a creamcolored solid (318 mg, 89% pure, yield: 55%).

LC-MS: Rt=0.77 min; MS m/z [M+H]⁺ 574.4/576.4, m/z [M−H]⁻ 572.3/574.3;UPLC-MS 1

LC-MS: Rt=3.90 min; MS m/z [M+H]⁺ 574.3/576.3, m/z [M−H]⁻ 572.3/574.2;UPLC-MS 2

Intermediate FA:2-(5-cyclopropyl-2-(3,6-dihydro-2H-pyran-4-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamideStep 1: 2-bromo-5-cyclopropyl-[1,2,4]triazolo[1,5-a]pyrimidin-7(4H)-one

5-Bromo-4H-1,2,4-triazol-3-amine (5.00 g, 29.1 mmol) and ethyl3-cyclopropyl-3-oxopropanoate (6.83 g, 43.7 mmol) were mixed in1-butanol (40 mL). H₃PO₄ (8.40 g, 72.9 mmol) was added and the RM wasstirred at 100° C. for 20 hours. Ethyl 3-cyclopropyl-3-oxopropanoate(1.00 g, 6.40 mmol) was added and the RM was stirred at 100° C. for 22.5hours. Ethyl 3-cyclopropyl-3-oxopropanoate (1.00 g, 6.40 mmol) was addedand the RM was stirred at 100° C. for 23.5 hours. The RM was cooled toRT and the yellow suspension was filtered. The cake was washed with asmall amount of EtOH and the filtrate was concentrated under reducedpressure. The cake was washed with hot EtOH and the filtrate wasconcentrated under reduced pressure. The cake and the filtrate containedproduct, so both were combined again and concentrated under reducedpressure. The resulting oil was stood at RT over the weekend. A solidcrystallized out which was filtered off and washed with Et₂O to give awhite solid (2.62 g). The crude product was adsorbed onto Isolute andpurified by column chromatography (Silica gel column: Silica 40 g,eluent DCM:MeOH 100:0 to 85:15). The product containing fractions werecombined and concentrated under reduced pressure to give the titlecompound as a white solid (923 mg, 99% pure, yield: 12%). The motherliquid was concentrated, adsorbed onto Isolute and purified by columnchromatography (Silica gel column: Silica 120 g, eluent DCM:MeOH 100:0to 85:15). The product containing fractions were combined andconcentrated under reduced pressure to give the title compound as abeige solid (1.14 g, 99% pure, yield: 15%).

Total: 2.06 g, 99% pure, yield: 27%).

LC-MS: Rt=0.50 min; MS m/z [M+H]⁺ 255.0/257.0, m/z [M−H]⁻ 252.9/254.9;UPLC-MS 4

Step 2:2-(2-bromo-5-cyclopropyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

2-Bromo-5-cyclopropyl-[1,2,4]triazolo[1,5-a]pyrimidin-7(4H)-one (1.19 g,4.67 mmol) and 2-bromo-N-(4-(trifluoromethyl)phenyl)acetamide (1.73 g,5.84 mmol) were mixed in DMF (12 mL). DIPEA (2.50 mL, 14.0 mmol) wasadded and the RM was stirred at 65° C. for 6 hours, then at RTovernight. Water was added and the RM was continued stirring at RT. Aresin collapsed. The suspension was filtered and the cake was washedwith water. The cake (1.78 g) was suspended in DCM and MeOH and filteredoff. Then it was washed and dried under Hv to give the title compound asa beige solid (829 mg, 85% pure, yield: 33%).

LC-MS: Rt=1.03 min; MS m/z [M+H]⁺ 456.1/458.1, m/z [M−H]⁻ 453.9/455.9;UPLC-MS 4

Step 3:2-(5-cyclopropyl-2-(3,6-dihydro-2H-pyran-4-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

2-(2-Bromo-5-cyclopropyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide(1.34 g, 2.94 mmol),2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(926 mg, 4.41 mmol) and Pd X-Phos G3 (124 mg, 147 μmol) were mixed indioxane (15 mL) and 1M K₃PO₄ in water (8.81 mL, 8.81 mmol) was added.The RM was vacuumed and backfilled with argon several times, then it wasstirred at 90° C. for 1.5 hours. The RM was cooled to RT. The RM wasextracted with EtOAc (3×70 mL) and water (2×20 mL). The organic layerwas dried through a phase separator and concentrated under reducedpressure. The aqueous layer was a suspension which was filtered. Theaqueous layer was extracted three times with DCM, dried through a phaseseparator and concentrated under reduced pressure. All organics werecombined with the cake and were suspended in hot EtOH (500 mL). Then itwas filtered, and the cake was dissolved in warm ACN and Si-Thiol (2.00g) was added. The mixture was stirred at 45° C. for 5 minutes, then itwas filtered. The filtrate was concentrated under reduced pressure togive the title compound as a grey solid (650 mg, 99% pure, yield: 48%).The mother liquor was mixed with Si-Thiol (2.00 g) and stirred at 45° C.for 5 minutes, then it was filtered. The filtrate was concentrated underreduced pressure to give the title compound as a bright brown solid (610mg, 79% pure, yield: 36%).

Total: 1.26 g, 89% pure, yield: 84%.

LC-MS: Rt=0.98 min; MS m/z [M+H]⁺ 460.3, m/z [M−H]⁻ 458.3; UPLC-MS 4

Step 4:2-(6-bromo-5-cyclopropyl-2-(3,6-dihydro-2H-pyran-4-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

2-(5-Cyclopropyl-2-(3,6-dihydro-2H-pyran-4-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide(610 mg, 1.33 mmol) and NBS (295 mg, 1.66 mmol) were mixed in DMF (28mL). The RM was stirred at 60° C. for 5.5 hours, then it was stood at RTover the weekend. NBS (125 mg, 703 μmol) was added and the RM wasstirred at 60° C. for 5 hours. NBS (50 mg, 281 μmol) was added and theRM was stirred at 60° C. for 1.5 hours. Then it was cooled to RT andstood at RT overnight. The RM was diluted with DCM and aq sat NaHCO₃.Most of the DMF was removed under reduced pressure. The solid residuewas extracted with EtOAc (3×40 mL), water (2×20 mL) and brine (25 mL).The organic layer was dried through a phase separator and concentratedunder reduced pressure. The brown solid residue was mixed with hexaneand the suspension was filtered. The cake was mixed again with hexaneand filtered again. The cake was dried under HV to give the titlecompound as a bright brown solid (508 mg, 74% pure, yield: 53%).

LC-MS: Rt=1.04 min; MS m/z [M+H]⁺ 538.1/540.1, m/z [M−H]⁻ 536.2/538.2;UPLC-MS 4

Step 5:2-(5-cyclopropyl-2-(3,6-dihydro-2H-pyran-4-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

2-(6-bromo-5-cyclopropyl-2-(3,6-dihydro-2H-pyran-4-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide(548 mg, 743 μmol) and piperazine (2.50 g, 29.0 mmol) were mixed withDMSO (5 mL) and the RM was stirred at 140° C. under argon for 5 hours.The RM was cooled to RT and stood at RT overnight, then it was combinedwith another batch. The RM was extracted with EtOAc (3×80 mL), aq satNaHCO₃ (2×30 mL) and water (2×30 mL). The organic layer was washed with1N HCl (4×25 mL) and water (2×20 mL). The organic layer was concentrateda bit and extracted twice with 1N HCl. The combined aqueous layers werebasified with solid NaHCO₃ and extracted three times with EtOAc. Theorganic layer was dried through a phase separator and concentrated underreduced pressure. The solid residue was suspended in DCM and MeOH andfiltered. The cake was washed well with DCM and the filtrate wasconcentrated down to give a bright brown solid (254 mg). The crudeproduct was adsorbed onto Isolute and purified by column chromatography(Silica gel column: Silica 24 g, eluent DCM:MeOH/Et₃N (95/5) 90:10 to50:50). The product containing fractions were combined and concentratedunder reduced pressure to give the title compound as a solid (97.0 mg,87% pure, yield: 21%).

LC-MS: Rt=0.86 min; MS m/z [M+H]⁺ 544.3, m/z [M−H]⁻ 542.3; UPLC-MS 3

Intermediate FB:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-(2-hydroxyethyl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamideStep 1: tert-butyl4-(2-bromo-5-methyl-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

A yellow solution of tert-butyl4-(2-bromo-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(Intermediate BM) (1.00 g, 2.42 mmol) in THF (10 mL) and DMF (2 mL) wascooled in an ice bath to 0° C. Then NaH 60% in mineral oil (116 mg, 2.90mmol) was added carefully portionwise, gas evolution was observed. TheRM was stirred at 0° C. for 30 minutes. Then(2-(chloromethoxy)ethyl)trimethylsilane (498 μL, 2.81 mmol) was addeddropwise. The cooling bath was removed and the RM was allowed to warm toRT and stirred for 1 day. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM(10 mL) were added. The aqueous layer was washed with DCM (2×10 mL). Thecombined organic layers were dried through a phase separator andconcentrated under reduced pressure to give the title compound (3.50 g,38% pure, quantitative).

LC-MS: Rt=1.46 min; MS m/z [M+H]⁺ 443.3/445.3, m/z [M−H]⁻ 411.3/413.2;UPLC-MS 1

Step 2:2-(2-bromo-6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-5-yl)aceticacid

Tert-butyl4-(2-bromo-5-methyl-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(245 mg, 311 μmol) was dissolved in THE (3 mL) and NaHMDS 1M in THE (373μL, 373 μmol) was added dropwise. The RM was stirred at −78° C. for 1.5hours. Then CO₂ (collected from dry ice) was bubbled in. The reactionwas stirred at −78° C. for 20 minutes. Water (10 mL), aq 1M HCl (10 mL)and DCM (10 mL) were added. The aqueous layer was washed with DCM (2×10mL). The combined organic layers were dried through a phase separatorand concentrated under reduced pressure to give the title compound (1.91g, 60% pure, yield: 80%).

LC-MS: Rt=1.31 min; MS m/z [M+H−Boc]⁺ 487.2/489.2; UPLC-MS 1

Step 3: tert-butyl4-(2-bromo-5-(2-methoxy-2-oxoethyl)-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

2-(2-Bromo-6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-5-yl)aceticacid (1.90 g, 2.01 mmol) was dissolved in THE (10 mL) and MeOH (2 mL)and cooled to 0° C. Then (diazomethyl)trimethylsilane 2M in Et₂O (1.10mL, 2.21 mmol) was added dropwise. The reaction was stirred at RT for 30minutes. Water (50 mL), aq sat NaHCO₃ (50 mL) and DCM (50 mL) wereadded. The aqueous layer was washed with DCM (2×50 mL). The combinedorganic layers were dried through a phase separator and concentratedunder reduced pressure to give the title compound (1.86 g, 67% pure,quantitative).

LC-MS: Rt=1.40 min; MS m/z [M+H−Boc]⁺ 501.3/503.3, m/z [M−H]⁻599.1/601.1; UPLC-MS 1

Step 4: tert-butyl4-(2-bromo-5-(2-methoxy-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-(2-methoxy-2-oxoethyl)-7-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(1.36 g, 1.99 mmol) was dissolved in DCM (5 mL) and TFA (3.00 mL, 13.0mmol) was added. The mixture was stirred at 40° C. for 2 hours. The RMwas concentrated under reduced pressure. The residue was suspended inDCM (5 mL) and DIPEA (696 μL, 3.99 mmol) was added, followed by Boc₂O(463 μL, 1.99 mmol). The mixture was stirred at RT for 1 hour. Water (10mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL) were added. The aqueous layerwas washed with DCM (2×10 mL). The combined organic layers were driedthrough a phase separator and concentrated under reduced pressure andconcentrated under reduced pressure to give the title compound (400 mg,35% pure, yield: 15%).

LC-MS: Rt=0.81 min; MS m/z [M+H]⁺ 471.2/473.2, m/z [M−H]⁻ 469.2/471.2;UPLC-MS 1

Step 5: tert-butyl4-(2-bromo-5-(2-hydroxyethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-(2-methoxy-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(100 mg, 74.0 μmol) was dissolved in THF (1 mL) and LiAlH₄ (111 μL, 223μmol) was added, the mixture was stirred at RT for 10 minutes. Water (10mL), aq sat NaHCO₃ (10 mL) and DCM (10 mL) were added. The aqueous layerwas washed with DCM (2×10 mL). The combined organic layers were driedthrough a phase separator and concentrated under reduced pressure togive the title compound (208 mg, 44% pure, yield: 70%).

LC-MS: Rt=0.76 min; MS m/z [M+H]⁺ 443.1/445.1, m/z [M−H]⁻ 441.3/443.1;UPLC-MS 1

Step 6: tert-butyl4-(2-bromo-5-(2-hydroxyethyl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-(2-hydroxyethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(208 mg, 206 μmol) and2-bromo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (IntermediateDP) (67.2 mg, 227 μmol) were mixed in dioxane (2 mL) and DIPEA (72.0 μL,413 μmol) was added. The mixture was stirred at 80° C. for 3 hours. Thenat 50° C. for 2 hours and then at 80° C. overnight. Water (10 mL), aqsat NaHCO₃ (10 mL) and DCM (10 mL) were added. The aqueous layer waswashed with DCM (2×10 mL). The combined organic layers were driedthrough a phase separator and concentrated under reduced pressure. Thecrude product was purified by reverse phase preparative HPLC (RP-HPLCacidic 1: 5 to 100% B in 20 min). The product containing fractions werecombined, basified with aq sat NaHCO₃, extracted twice with DCM, driedthrough a phase separator and concentrated under reduced pressure togive the title compound (45.4 mg, 82% pure, yield: 27%).

LC-MS: Rt=1.23 min; MS m/z [M+H−Boc]⁺ 558.2/560.2, m/z [M−H]⁻656.5/658.5; UPLC-MS 1

Step 7: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-(2-hydroxyethyl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate

Tert-butyl4-(2-bromo-5-(2-hydroxyethyl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(264 mg, 388 μmol),2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(136 mg, 647 μmol) and Pd X-Phos G3 (16.4 mg, 19.0 μmol) were mixed indioxane (2 mL) and K₃PO₄ 1M in water (1.16 mL, 1.16 mmol) was added. Themixture was stirred at 80° C. for 20 minutes. Water (10 mL), aq satNaHCO₃ (10 mL) and DCM (10 mL) were added. The aqueous layer was washedwith DCM (2×10 mL). The combined organic layers were dried through aphase separator and concentrated under reduced pressure. The crudeproduct was redissolved in DCM and MeOH and SiliaMetS® Thiol was added.The mixture was stirred at 40° C. for 1 hour. Then it was filtered. Thecake was washed with DCM. The filtrate was concentrated under reducedpressure to give the title compound (905 mg, 86% pure).

LC-MS: Rt=1.20 min; MS m/z [M+H]⁺ 662.3, m/z [M−H]⁻ 660.5; UPLC-MS 1

Step 8:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-(2-hydroxyethyl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

Tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-(2-hydroxyethyl)-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate(925 mg, 318 μmol) was dissolved in DCM (5 mL) and TFA (1.00 mL, 13.0mmol) was added. The mixture was stirred at 40° C. for 3 hours. Theproduct seems not to be stable under acidic conditions at 40° C. so thereaction was stopped. Water (10 mL), aq sat NaHCO₃ (10 mL) and DCM (10mL) were added. The aqueous layer was washed with DCM (2×10 mL). Thecombined organic layers were dried through a phase separator andconcentrated under reduced pressure to give the title compound (168 mg,28% pure, yield: 26%).

LC-MS: Rt=1.20 min; MS m/z [M+H]⁺ 562.2, m/z [M−H]⁻ 560.4; UPLC-MS 1

Intermediate FC:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-6-(piperidin-4-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamideStep 1: 2-bromo-5-methyl-[1,2,4]triazolo[1,5-a]pyrimidin-7(4H)-one

To a solution of 3-bromo-1H-1,2,4-triazol-5-amine (Intermediate ER)(60.0 g, 60.6 mmol) in AcOH (380 mL) was added ethyl 3-oxobutanoate(86.6 g, 66.6 mmol). The RM was stirred at 80° C. overnight. The mixturewas filtrate and washed with AcOH (160 mL). The wet filter cake wasdried to give the title compound (80.0 g, 80%).

Step 2:2-bromo-6-iodo-5-methyl-[1,2,4]triazolo[1,5-a]pyrimidin-7(4H)-one

2-Bromo-5-methyl-[1,2,4]triazolo[1,5-a]pyrimidin-7(4H)-one (2.00 g, 8.73μmol) was added in a flame dried flask under nitrogen. Acetic acid (29.1mL) was added, followed by NIS (2.16 g, 9.61 mmol). The RM was stirredat 60° C. for 1 hour. The RM was cooled to RT. The solid was filteredoff and washed 3 times with EtOH. The solid was dried under Hv to givethe title compound (2.76 g, 95% pure, yield: 89%).

LC-MS: Rt=0.56 min; MS m/z [M+H]⁺ 354.9/356.9, m/z [M−H]⁻ 353.0/355.0;UPLC-MS 8

Step 3: tert-butyl4-(2-bromo-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,6-dihydropyridine-1(2H)-carboxylate

Palladium G3-Tricyclohexylphosphine,[(Tricyclohexylphosphine)-2-(2-aminobiphenyl)]palladium(II)methanesulfonate (183 mg, 282 μmol) was purged with nitrogen. A solutionof 2-bromo-6-iodo-5-methyl-[1,2,4]triazolo[1,5-a]pyrimidin-7(4H)-one(1.00 g, 2.82 mmol) and tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate(894 mg, 2.89 mmol) in n-butanol (100 μL) was added, followed by K₃PO₄1.5M in water (5.63 mL, 8.45 mmol). The RM was stirred at 70° C. for 1hour. Tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate(50.0 mg, 162 μmol) was added and the RM was stirred at 70° C.overnight. N-butanol was removed under reduced pressure. EtOAc was addedand the mixture was washed with NH₄Cl. The organic layer was driedthrough a phase separator. ISOLUTE® Si-TMT (6.50 g, 2.82 mmol) was addedand the mixture was stirred at 40° C. for 1 hour. The solid was filteredoff and washed with EtOAc. The filtrate was concentrated under reducedpressure. The crude product was purified by column chromatography(Silica gel column: Silica 40 g, eluent DCM:DCM/MeOH (8/2) 100:0 to50:50) to give the title compound as a white powder (734 mg, 90% pure,yield: 57%).

LC-MS: Rt=0.86 min; MS m/z [M−H]⁻ 408.2/410.2; UPLC-MS 14

Step 4: tert-butyl4-(2-bromo-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperidine-1-carboxylate

To a solution of tert-butyl4-(2-bromo-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,6-dihydropyridine-1(2H)-carboxylate(734 mg, 1.79 mmol) in MeOH (10 mL) was added PtO₂ (40.6 mg, 179 μmol)under N2 atmosphere. The flask was purged with hydrogen for 2 minutes.The RM was stirred at RT for 2 hours. PtO₂ (66.0 mg, 291 μmol) was addedand the RM was stirred at RT for 2 hours. PtO₂ (103 mg, 454 μmol) wasadded and the RM was stirred at RT overnight. PtO₂ (122 mg, 543 μmol)was added and the RM was stirred at RT. The RM was filtered through apad of celite. The crude product was purified by column chromatography(Silica gel column: Silica 12 g, eluent DCM:DCM/MeOH (8/2) 100:0 to50:50) to give the title compound (580 mg, 80% pure, yield: 63%).

LC-MS: Rt=0.89 min; MS m/z [M+H]⁺ 412.2/414.2, m/z [M−H]⁻ 410.3/412.3;UPLC-MS 8

Step 5: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperidine-1-carboxylate

Tert-butyl4-(2-bromo-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperidine-1-carboxylate(580 mg, 1.41 mmol),2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(443 mg, 2.11 mmol) and X-Phos Pd G3 (71.4 mg, 84.0 μmol) were mixed.Under N2 atmosphere were added DMF (1.4 mL) and K₃PO₄ 1M in water (2.81mL, 1.82 mmol). The RM was stirred at 80° C. for 1 hour. X-Phos Pd G3(10.0 mg, 11.8 μmol) and2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(60.0 mg, 286 μmol) were added and the RM was stirred for 3 hours togive the title compound as RM.

LC-MS: Rt=1.25 min; MS m/z [M+H]⁺ 416.3, m/z [M−H]⁻ 414.4; UPLC-MS 8

Step 6: tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperidine-1-carboxylate

To the RM containing tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperidine-1-carboxylatewas added 2-bromo-N-(4-(trifluoromethyl)phenyl)acetamide (397 mg, 1.41mmol). The RM was stirred at 80° C. for 40 minutes.2-Bromo-N-(4-(trifluoromethyl)phenyl)acetamide (88.0 mg, 313 μmol) wasadded and the RM was stirred at 80° C. for 30 minutes. Most of the DMFwas removed under reduced pressure. EtOAc was added and the mixture waswashed with NaHCO₃. The organic layer was dried through a phaseseparator and concentrated under reduced pressure. The mixture wastreated with Si TMT. The solvent was removed, the residue was adsorbedonto Isolute and purified by column chromatography (Silica gel column:Silica 24 g, eluent DCM:DCM/MeOH (8/2) 100:0 to 70:30), then in 5portions by reverse phase preparative HPLC (5×RP-HPLC acidic 1: 20 to95% B in 20 min), to give the title compound (185 mg, 60% pure, yield:18%).

LC-MS: Rt=1.18 min; MS m/z [M+H]⁺ 617.4, m/z [M−H]⁻ 615.5; UPLC-MS 8

Step 7:2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-6-(piperidin-4-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)phenyl)acetamide

To tert-butyl4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)phenyl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperidine-1-carboxylate(145 mg, 198 μmol) was added TFA (306 μL, 3.97 mmol) in DCM (2 mL). TheRM was stirred at RT for 1 hour. DCM was added and the crude product waswashed with NaOH solution. The aqueous layer was washed with EtOAc. Theorganic layer was dried through a phase separator and concentrated underreduced pressure to give the title compound (130 mg, 79% pure,quantitative).

LC-MS: Rt=0.74 min; MS m/z [M+H]⁺ 517.3, m/z [M−H]⁻ 515.4; UPLC-MS 8

Synthetic Schemes for the Compounds of Formulae 1b, 1c, 1d, 1e and 1f

Processes are provided to make to compounds of formulae 1b, 1c, 1d, 1eand 1f. Unless otherwise stated, the groups of the process schemes areas defined in the embodiments and preferences herein. The syntheses canbe modified to make variants under formula (I), according to proceduresknown to the skilled chemist.

A process is provided for preparing a compound of formula AAK (SchemeXI) comprising steps a, b, c, d, e, f, g, h, i, and j. It is understoodthat the order of process steps a, b, c, d, e, f, g, h, i and j may bechanged to optimize the synthesis as necessary. The compound of formulaAAK can be obtained via coupling reaction step j by reacting compound offormula AAJ with compound AAZ wherein R₄ is defined above. The couplingreaction can be an amide formation. The coupling reaction step can becarried out with for example HATU((1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxide hexafluorophosphate) or alternatively Ghosez reagent(1-Chloro-N,N,2-trimethylpropenylamine), preferably in a one or two stepprocedure. Alternatively, further alternative amide coupling methods areknown in the art. For examples of amide bond formations, seeMantalbetti, C. A. G. N and Falque, V., Amide bond formation and peptidecoupling, Tetrahedron, 2005, 61(46), pp 10827-10852 and references citedtherein.

Compound of formula AAJ can be prepared comprising step i ofdeprotecting PG from the compound of formula AAI, wherein PG representsa suitable protecting group, preferably a BOC group, and wherein theother substituents are as defined above. There are many known ways ofdeprotecting BOC groups. The deprotection step can be carried out withfor example TFA or HCl in a solvent for example dichloromethane ordioxane.

Compound of formula AAI can be prepared comprising step h starting froma compound of formula AAH wherein R₅₀ represents halo, particularlybromo and wherein PG represents a suitable protecting group for examplea BOC group and the other substituents are as defined above. Step h canbe a nucleophilic aromatic substitution reaction and can be carried outby combining compound of formula AAH with an amine for exampletert-butyl piperazine-1-carboxylate or alternatively piperazine. Astoichiometric excess of the amine can be used, preferably between 2 and50 mole equivalents in an organic solvent for example DMSO or NMP. Thereaction is preferably stirred at a temperature of approximately 80-140°C. and can be carried out in a capped tube. An alternative method forstep h can use Buchwald-Hartwig conditions using a amine for exampletert-butyl piperazine-1-carboxylate, a ligand such as Brettphos orRuPhos or RuPhos hybrid with a palladium catalyst such as RuPhos Pd G1,RuPhos Pd G4 or [PdCl(allyl)]2 in the presence of a base such as K2CO3or Cs2CO3 or tert-BuONa in an organic solvent such as dioxane or THF.The reaction is preferably stirred at a temperature of approximately80-120° C. The reaction is preferably carried out under an inert gassuch as nitrogen or argon. Alternatively, further alternativeBuchwald-Hartwig coupling methods are known in the art, for examples ofmethods, see B. T. Ingoglia et al Biaryl monophosphine ligands inpalladium-catalyzed C—N coupling: An updated User's guide, Tetrahedron,2019, 75(32), pp 4199-4211 and references cited therein.

Compound of formula AAH can be prepared comprising step g whereincompound of formula AAG, wherein the substituents are defined as herein,is halogenated. Step g can be carried out using a halogenating reagentsuch as N-bromosuccinimide or N-iodosuccinimide in a solvent for exampleDMF or acetonitrile. The reaction is preferably stirred at a temperatureof approximately 20-80° C.

Compound of formula AAG can be prepared comprising step f whereincompound of formula AAF, wherein the substituents are defined as herein,is alkylated by reacting compound of formula AAY wherein R₅₀ representshalo, particularly bromo, iodo or chloro and the other substituents aredefined as above. Step f can be carried out in the presence of a basesuch as K₂CO₃ or N,N-diisopropylethylamine in a solvent for example DMFor dioxane. The reaction is preferably stirred at a temperature ofapproximately 20-80° C.

Compound of formula AAF can be prepared comprising step e starting fromcompound of formula AAE wherein R₅₁ represents H or methoxy and theother substituents are as defined above. Many methods of cleaving benzylor para-methoxybenzyl groups are known in the art. Step e can be carriedout in the presence of an acid such as TFA or HCl or HBr, preferably instoichiometric excess in a solvent such as dichloromethane or dioxaneand is preferably stirred at a temperature of approximately 20-80° C. Analternative method for step e can use hydrogenation conditions in thepresence of a hydrogen atmosphere and a catalyst such as Pd/C orpalladium hydroxide/C. The reaction is preferably stirred at atemperature of approximately 20-50° C. in an organic solvent such asethanol or methanol.

Compound of formula AAE can be prepared comprising step d starting fromcompound of formula AAD wherein R₅₀ represents halo, particularly bromoand the other substituents are as defined above. Step d comprisesreacting 2-10 mol equivalents of an alcohol for example benzyl alcoholor para-methoxybenzyl alcohol with 2-5 mol equivalents of a base such assodium hydride in an organic solvent such as THF or dioxane withstirring at a temperature of approximately 20-40° C., preferably 20° C.for approximately 10-60 minutes. A compound of formula AAD is then addedand stirring is continued at a temperature of approximately 20-100° C.,preferably 20-60° C. The reaction is preferably carried out under aninert gas such as nitrogen or argon. An example method is described inWO2021/222522, 2021, A1 page 574.

Compound of formula AAD wherein R₅₀ represents halo, particularly bromoor iodo, and the other substituents are as defined herein, can beprepared comprising step c starting from compound of formula AAC. Step ccomprises reacting a compound of formula AAC with a base such as LiTMP(lithium tetramethylpiperidide) or LDA (lithium diisopropylamide) withstirring in a solvent such as THF at a temperature of approximately −78°C. to 20° C. under an inert gas such as nitrogen or argon. Afterstirring for an appropriate time, approximately 30 minutes to 3 hours, ahalogenating reagent such as bromine or iodine is then added at atemperature of approximately −78° C. to 20° C. and stirring iscontinued. Other suitable halogenating agents are known in the art.

Compound of formula AAC wherein R₁ and R₃ are as defined above can beprepared comprising step b starting from compound of formula AAB. Step bcan be a Suzuki or Negishi or Stille or Kumada cross-coupling reactionand comprises reacting a compound of formula AAB with R₃n-MX wherein R₃is as defined above, n is 1,2,3 or 4 and MX represents for exampleB(OH)2, BPin (Pin represents boronic acid pinacol ester) BF3K, B(MIDA),Sn, Zn, Mg-Halo. Example Negishi cross-coupling conditions comprisereacting compound of formula AAD with an alkyl zincate for exampledimethylzinc or diethylzinc, preferably in stoichiometric excess forexample 2-10 mol equivalents, in the presence of a catalyst such asPdCl2(dppf) or Pd(PPh₃)₄ in a suitable solvent such as THF at atemperature of approximately 20-120° C., preferably 20-80° C. under aninert gas such as nitrogen or argon.

Compound of formula AAB wherein R₁ is as defined above can be preparedcomprising step a from compound AAA. Step a can be a Suzuki or Negishior Stille or Kumada cross-coupling reaction and comprises reacting acompound of formula AAA with R1 n-MX wherein R₁ is as defined above, nis 1,2,3 or 4 and MX represents for example B(OH)₂, BPin (Pin representsboronic acid pinacol ester) BF₃K, Sn, Zn, Mg-Halo. Example Suzukicross-coupling conditions comprise reacting compound of formula AAA withR₁-BPin in the presence of a catalyst such as PdCl₂(dppf) or Pd(PPh₃)₄and a base such as K₃PO₄ or potassium carbonate in a suitable solventmixture such as DMF, THF or dioxane or water at a temperature ofapproximately 20-120° C., preferably 20-80° C. under an inert gas suchas nitrogen or argon. An example method is described in CN112707908 Apage 32.

Compound of formula AAA can be prepared according to the methoddescribed in CN112707908 A page 31.

Alternatively compound of formula AAK can be prepared according to theroute shown in Scheme XII comprising steps k, L, m, zd, za, zh, ze, zjand zf. Methods comprising steps zd, za, zh, ze, zj and zf to preparecompounds of formulas AAP, AAQ, AAR, AAS and AAT can be performed usinganalogous conditions as those described above for steps d, a, h, e, jand f for Scheme XI. It is understood that the order of process steps k,L, m, zd, za, zh, ze, zj and zf may be changed to optimize the synthesisas necessary.

Compound of formula AAO wherein R₅₀ represents halo, particularly bromoor chloro, and the other substituents are as defined above, can beprepared comprising step m starting from compound of formula AAN. Step mcomprises reacting a compound of formula AAN with a halogenating agentsuch as PCl₅ or PBr₃ in stoichiometric excess for example 2-10 molequivalents in a sealed tube at a temperature of approximately 200-270°C., preferably 250-270° C. for approximately 1-10 hours. An examplemethod is described in J. Org. Chem., Vol. 39, No. 15,1974, page 2146.

Compound of formula AAN wherein R₃ is as defined above, can be preparedcomprising step L starting from compound of formula AAM. Step Lcomprises reacting a compound of formula AAM with hydroxylamine in thepresence of a base such as triethylamine and a solvent such as ethanolor methanol at a temperature of approximately 60-100° C. The product ofthis reaction is then reacted with tert-butyl nitrite in the presence ofCuBr₂ in a solvent such as acetonitrile at a temperature ofapproximately 20-50° C. An example method is described in CN112707908 Apage 31.

Compound of formula AAM wherein R₃ is as defined above, can be preparedcomprising step k starting from AAL. Step k comprises reacting acompound of formula AAL with ethoxycarbonyl isothiocyanate in a solventsuch as dichloromethane at a temperature of approximately 0-20° C. for2-18 hours.

Compound of formula AAL wherein R₃ is as defined above are commerciallyavailable, or methods for their preparation are known in the art.

Compound of formula 1d can be prepared according to the example routeshown in Scheme XIII using analogous methods to those described herein.Analogous methods to chemists skilled in the art can be adaptedaccordingly. It is understood that the order of process steps shown inScheme XIII may change to optimize the synthesis as necessary.

Compound of formula 1e can be prepared according to the example routeshown in Scheme XIV using analogous methods to those described herein.Analogous methods to chemists skilled in the art can be adaptedaccordingly. It is understood that the order of process steps shown inScheme XIV may change to optimize the synthesis as necessary.

A process for preparing compound of formula BBN (Scheme XV) comprisingsteps ba, bc, bd, be, bf, bg, bh, bj, bk, bL, bm, ye or yi and yj. It isunderstood that the order of process ba, bc, bd, be, bf, bg, bh, bj, bk,bL, bm, ye or yi and yj may change to optimize the synthesis asnecessary. The compound of formula BBN can be obtained via couplingreaction step yj by reacting compound of formula BBM wherein thesubstituents are as defined above with compound AAZ wherein R₄ is asdefined above using analogous methods to those described herein.

Compound of formula BBM wherein the substituents are as defined abovecan be prepared deprotecting compound of formula BBL wherein PGrepresents a suitable protecting group such as BOC or para-methoxybenzylor benzyl and the other substituents are as defined above comprisingstep ye or step yi using analogous methods to those described for step eor step i for Scheme XI.

Compound of formula BBL can be prepared comprising step bL starting fromcompound BBK wherein the substituents are as defined above with eithercompound BBX or compound BBW wherein PG is as defined above and LG isrepresented by halo, particularly iodo or bromo or OH or OMs(methanesulfonate) or OTs (p-toluenesulfonate) or OTf(trifluoromethanesulfonate) or B(OH)2, BPin (Pin represents boronic acidpinacol ester) BF3K. Step bL can be performed by combining compound offormula BBK with compound BBX in the presence of a base such as sodiumhydride or K2CO3 or DBU or NaOtBu or phosphazene base P2-Et. Astoichiometric excess BBX can be used, preferably between 2 and 50 moleequivalents in an organic solvent for example DMF or NMP. The reactionis preferably stirred at a temperature of approximately 80-150° C. andcan be carried out in a capped tube. An alternative method for step bLcan use Ullmann-type reaction. Example Ullmann-type cross-couplingconditions comprise reacting compound of formula BBK with compound BBWin the presence of a catalyst such as copper(I)iodide, a ligand such asN-(2-cyanophenyl)pyridine-2-carboxamide or4,7-dimethoxy-1,10-phenanthroline or N1,N2-dibenzylethane-1,2-diamineand a base such as K3PO4 or K2CO3 in a suitable solvent mixture such asDMSO or DMF at an approximate temperature of 80-150° C. A stoichiometricexcess BBX can be used, preferably between 2 and 50 mole equivalents. Analternative method for step bL can comprise reacting compound of formulaBBK with compound BBW using Buchwald-Hartwig conditions using forexample using an analogous method to those described for step h (SchemeXI). The product of the reaction between the compound of formula BBK andthe compound of formula BBW can optionally be hydrogenated using methodsknown in the art, to give a compound of formula BBL wherein thepiperidine ring is saturated. Alternative cross-coupling conditions areknown in the art, for examples of methods, see De Meijere et aL.Metal-Catalyzed Cross-Coupling Reactions, Wiley, 2014 and referencescited therein.

Compound of formula BBK can be prepared comprising step bk starting fromcompound of formula BBJ and the substituents are as defined above. Stepbk comprises reacting a compound of formula BBJ with a stoichiometricexcess of L-methionine for example 3 to 5 mol equivalents in a solventsuch as methanesulfonic acid at approximate temperature of 20-80° C.

Compound of formula BBJ can be prepared comprising step bj starting fromcompound of formula BBI wherein the substituents are as defined above.Step bj comprises reacting a compound of formula BBI with astoichiometric excess of R₂—NH2 wherein R₂ is as defined above forexample 3 to 5 mol equivalents in the presence of a stoichiometricexcess of trimethylaluminium for example 3-5 mol equivalents in asolvent such as toluene at approximate temperature of 20-80° C.

Compound of formula BBI can be prepared comprising step bi starting fromcompound of formula BBH wherein the substituents are as defined above.Step bj comprises reacting a compound of formula BBH with astoichiometric excess of gaseous hydrogen chloride for example 20-100mol equivalents in ethanol at approximate temperature of 20-100° C.preferably in a sealed tube.

Compound of formula BBH can be prepared comprising step bh starting fromcompound of formula BBG wherein the substituents are as defined above.Step bh comprises reacting a compound of formula BBH with astoichiometric excess of potassium fluoride for example 3 to 5 molequivalents in water in the presence of an additional solvent such asDMF or methanol at an approximate temperature of 20-100° C. preferably60-100° C.

Compound of formula BBG can be prepared comprising step bg starting fromcompound of formula BBF wherein R₅₀ is represented by halo, particularlyiodo or bromo and the other substituents are as defined above. Step bgcomprises reacting a compound of formula BBF with4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoxazole in the presenceof a catalyst such as XPhos Pd G3 and a base such as potassium carbonatein a suitable solvent such as DMF at a temperature of approximately20-120° C., preferably 60-100° C. under an inert gas such as nitrogen orargon.

Compound of formula BBF can be prepared comprising step bf starting fromcompound of formula BBE wherein the substituents are as defined above.Step bf comprises reacting a compound of formula BBE with a base such assodium hydride at an approximate temperature of 0-20° C. in a solventsuch as DMF for approximately 5 to 30 minutes under an inert gas such asnitrogen or argon. 1-(bromomethyl)-4-methoxybenzene is then added andthe reaction is stirred at a temperature of approximately 0-20° C.

Compound of formula BBE can be prepared comprising step be whereincompound of formula BBD wherein the substituents are defined as above ishalogenated. Step be can be carried out using a halogenating reagentsuch as N-bromosuccinimide or N-iodosuccinimide in a solvent for exampleDMF or acetonitrile. The reaction is preferably stirred at a temperatureof approximately 20-80° C.

Compound of formula BBD can be prepared comprising step bd starting fromcompound of formula BBC wherein the substituents are as defined above.Step bd comprises reacting a compound of formula BBC with a compound offormula BBY wherein R1 is defined as above in a solvent such as DMF ortoluene or dioxane at a temperature of approximately 80-150° C. Analternative method for step bd can comprise reacting compound of formulaBBC with a compound of formula BBZ wherein R1 is defined as above in asolvent such as dichloroethane or DMF or toluene or dioxane at atemperature of approximately 0-20° C. A base such as triethylamine maybe added. The reaction is then stirred at an approximate temperature of80-150° C.

Compound of formula BBC can be prepared comprising step bc starting fromcompound of formula BBB wherein the substituents are defined as above.Step bc comprises reacting a compound of formula BBB with astoichiometric excess of hydrazine hydrate for example 2 to 5 molequivalents in a solvent such as ethanol. The reaction is preferablystirred at a temperature of approximately 60-100° C.

Compound of formula BBB can be prepared comprising step ba starting fromcompound of formula BBA or BBAA wherein the substituents are defined asabove. Compound of formula BBA or BBAA are commercially available, ormethods for their preparation are known in the art. Step ba comprisesreacting a compound of formula BBA or BBAA with P2S5 or lawessonsreagent in a solvent such as dioxane or pyridine. The reaction ispreferably stirred at a temperature of approximately 80-120° C.

Compound of formula CCN can be prepared according to the route shown inScheme XVI comprising steps ca, cb, cc, cd, ce, cf, xg, xh, xi, xj, xk,xe or xi and xj. It is understood that the order of process ca, cb, cc,cd, ce, cf, xg, xh, xi, xj, xk, xe or xi and xj may change to optimizethe synthesis as necessary. Methods comprising steps xg, xh, xi, xj, xk,xe or xi and xj to prepare compounds of formulas CCG, CCH, CCl, CCJ,CCL, CCM and CCN can be performed using analogous conditions as thosedescribed above for steps bf, bg, bh, bi, e, i and j for Scheme XI andScheme XV.

Compound of formula CCK can be prepared comprising step xj starting fromcompound of formula CCJ wherein the substituents are defined as above.Step xj comprises reacting a compound of formula CCJ with di-tert-butyldicarbonate or para-methoxybenzyl bromide or benzyl bromide in thepresence of a base such as triethylamine in a solvent such asdichloromethane or dioxane at a temperature of approximately 0-20° C.

Compound of formula CCF wherein PG represents a suitable protectinggroup such as BOC or para-methoxybenzyl or benzyl and the othersubstituents are as defined above can be prepared comprising step cestarting from compound of formula CCE wherein substituents are asdefined above. Step ce comprises reacting a compound of formula CCE withEchavarren's gold(I) catalyst in a solvent such as THF at a temperatureof approximately 60-140° C., preferably 80-120° C. in a sealed tube. Anexample method is described in Org. Lett. 2013, 15, 11, 2616-2619. Analternative method of preparing compound of formula CCF comprisesreacting compound of formula CCE with a base such as sodium hydride in asolvent such as DMF or THF or dioxane at a temperature of approximately60-140° C., preferably 80-120° C. under an inert gas such as nitrogen orargon in a sealed tube.

Compound of formula CCE can be prepared comprising step cd starting fromcompound CCD wherein the substituents are defined as above. Step cdcomprises reacting a compound of formula CCD with compound of formulaCCY or CCZ wherein PG represents a suitable protecting group such as BOCor para-methoxybenzyl or benzyl in the presence of a base such astriethylamine or N-ethyl-N,N-diisopropylamine in a solvent such as THFor dioxane or DMF at a temperature of approximately 20-140° C.preferably 60-120° C. Compound of formula CCY or CCZ are commerciallyavailable, or methods for their preparation are known in the art.

Compound of formula CCD can be prepared comprising step cc starting fromcompound CCC wherein PG2 represents a protecting group such as MOM(methoxymethyl) or SEM (trimethylsilyl)ethoxymethyl) and the othersubstituents are as defined above. Step cc comprises reacting a compoundof formula CCC with an acid such as HCl or TFA in a solvent such asdioxane or dichloromethane at a temperature of approximately 0-80° C.preferably 20-60° C. Alternative methods for deprotection of SEM or MOMgroups are known in the art.

Compound of formula CCC can be prepared comprising step cb starting fromcompound of formula CCB wherein the substituents are as defined above.Step cb can be a Sonogashira reaction reacting a compound of formula CCBwith a compound of formula CCX wherein R₃ is as defined above in thepresence of a catalyst such as Pd(PPh₃)₄ and a copper catalyst such ascopper(I) iodide and a base such as triethylamine or lithium carbonatein a solvent such as dioxane or DMF or acetonitrile THF at a temperatureof approximately 20-120° C., preferably 80-120° C. under an inert gassuch as nitrogen or argon. Step cb can alternatively be a Suzuki orStille cross-coupling reaction and comprises reacting a compound offormula CCB with a compound of formula CCW wherein MX2 representsB(OH)₂, BPin (Pin represents boronic acid pinacol ester) BF₃K, B(MIDA),tributyltin and R₃ is as defined above. Methods for Sonogashira, Suzukior Stille reactions are known in the art. for examples of methods, seeMolnar et al. Palladium-Catalyzed Coupling Reactions, Wiley, 2013 andreferences cited therein.

Compound of formula CCB can be prepared comprising step ca starting fromcompound of formula CCA wherein the substituents are defined as above.Step a can be a Suzuki cross-coupling reaction and comprises reacting acompound of formula CCA with R_(1n)-MX wherein R1 is as defined above, nis 1,2,3 or 4 and MX represents for example B(OH)₂, BPin (Pin representsboronic acid pinacol ester) BF₃K. Example Suzuki cross-couplingconditions comprise reacting compound of formula CCA with R₁-BPin in thepresence of a catalyst such as PdCl₂(dppf) or Pd(PPh₃)₄ and a base suchas K₃PO₄ or potassium carbonate in a suitable solvent mixture such asDMF, THE or dioxane or water at a temperature of approximately 20-120°C., preferably 60-120° C. under an inert gas such as nitrogen or argon.Compound of formula CCA are commercially available, or methods for theirpreparation are known in the art.

Compound of formula DDN can be prepared according to the route shown inScheme XVII comprising steps da, db, dc, dd, de, df, dg, dh, di, dj, dk,dL or dM and dn. It is understood that the order of process da, db, dc,dd, de, df, dg, dh, di, dj, dk, dL or dM and dn may change to optimizethe synthesis as necessary. Methods comprising steps de, df, dg, dh, di,dj, dk, dL, dM and dn to prepare compounds of formulas DDF, DDG, DDH,DDI, DDJ, DDK, DDL, DDM and DDN can be performed using analogousconditions as those described herein, including optional hydrogenationsteps, for example step dk.

Compound of formula DDE wherein R₅₀ represents halo particularly bromoor iodo and the other substituents are as defined above can be preparedcomprising step dd starting from compound of formula DDD wherein thesubstituents are as defined above. Step dd can be carried out using ahalogenating reagent such as N-bromosuccinimide or N-iodosuccinimide ina solvent for example DMF or acetonitrile or acetic acid. The reactionis preferably stirred at a temperature of approximately 20-110° C.

Compound of formula DDD wherein the substituents are as defined abovecan be prepared comprising step dc starting from compound of formula DDCwherein the substituents are as defined above. Step dc can be performedusing analogous conditions as those described above for step a forScheme XI.

Compound of formula DDC wherein the substituents are as defined abovecan be prepared comprising step db starting from compound of formula DDBwherein the substituents are as defined above. Step db comprisesreacting compound of formula DDB with a stoichiometric excess of anammonium salt such as ammonium acetate for example 3 to 100 molequivalents particularly 10 to 20 mol equivalents in a solvent such asacetic acid at an approximate temperature of 60-130° C., preferably80-120° C.

Compound of formula DDB wherein the substituents are as defined abovecan be prepared comprising step da starting from compound of formulaDDA. Step da comprises reacting compound of formula DDA with compound offormula DDZ wherein R₅₀ represents halo particularly chloro or bromo andthe other the substituents are as defined above in the presence of abase such as potassium carbonate in a solvent such as acetone oracetonitrile at an approximate temperature of 0-50° C., preferably 0-20°C. Compound of formula DDZ are commercially available, or methods fortheir preparation are known in the art.

“Protecting Group”:

In the methods describe above, functional groups which are present inthe starting materials and are not intended to take part in thereaction, are present in protected form if necessary, and protectinggroups that are present are cleaved, whereby said starting compounds mayalso exist in the form of salts provided that a salt-forming group ispresent and a reaction in salt form is possible. In additional processsteps, carried out as desired, functional groups of the startingcompounds which should not take part in the reaction may be present inunprotected form or may be protected for example by one or moreprotecting groups. The protecting groups are then wholly or partlyremoved according to one of the known methods. Protecting groups, andthe manner in which they are introduced and removed are described, forexample, in “Protective Groups in Organic Chemistry”, Plenum Press,London, N.Y. 1973, and in “Methoden der organischen Chemie”,Houben-Weyl, 4th edition, Vol. 15/1, Georg-Thieme-Verlag, Stuttgart 1974and in Theodora W. Greene, “Protective Groups in Organic Synthesis”,John Wiley & Sons, New York 1981. A characteristic of protecting groupsis that they can be removed readily, i.e. without the occurrence ofundesired secondary reactions, for example by solvolysis, reduction,photolysis or alternatively under physiological conditions

The invention further includes any variant of the present processes, inwhich an intermediate product obtainable at any stage thereof is used asstarting material and the remaining steps are carried out, or in whichthe starting materials are formed in situ under the reaction conditions,or in which the reaction components are used in the form of their saltsor optically pure antipodes.

Compounds of the invention and intermediates can also be converted intoeach other according to methods generally known to those skilled in theart.

Intermediates and final products can be worked up and/or purifiedaccording to standard methods, e.g. using chromatographic methods,distribution methods, (re-) crystallization, and the like.

The following applies in general to all processes mentioned hereinbefore and hereinafter. All the above-mentioned process steps can becarried out under reaction conditions that are known to those skilled inthe art, including those mentioned specifically, in the absence or,customarily, in the presence of solvents or diluents, including, forexample, solvents or diluents that are inert towards the reagents usedand dissolve them, in the absence or presence of catalysts, condensationor neutralizing agents, for example ion exchangers, such as cationexchangers, e.g. in the H+ form, depending on the nature of the reactionand/or of the reactants at reduced, normal or elevated temperature, forexample in a temperature range of from about −100° C. to about 190° C.,including, for example, from approximately −80° C. to approximately 150°C., for example at from −80 to −60° C., at room temperature, at from −20to 40° C. or at reflux temperature, under atmospheric pressure or in aclosed vessel, where appropriate under pressure, and/or in an inertatmosphere, for example under an argon or nitrogen atmosphere.

At all stages of the reactions, mixtures of isomers that are formed canbe separated into the individual isomers, for example diastereoisomersor enantiomers, or into any desired mixtures of isomers, for exampleracemates or mixtures of diastereoisomers, for example analogously tothe methods described herein above.

The solvents from which those solvents that are suitable for anyparticular reaction may be selected include those mentioned specificallyor, for example, water, esters, such as lower alkyl-lower alkanoates,for example ethyl acetate, ethers, such as aliphatic ethers, for examplediethyl ether, or cyclic ethers, for example tetrahydrofuran or dioxane,liquid aromatic hydrocarbons, such as benzene or toluene, alcohols, suchas methanol, ethanol or 1- or 2-propanol, nitriles, such asacetonitrile, halogenated hydrocarbons, such as methylene chloride orchloroform, acid amides, such as dimethylformamide or dimethylacetamide, bases, such as heterocyclic nitrogen bases, for examplepyridine or N-methylpyrrolidin-2-one, carboxylic acid anhydrides, suchas lower alkanoic acid anhydrides, for example acetic anhydride, cyclic,linear or branched hydrocarbons, such as cyclohexane, hexane orisopentane, methycyclohexane, or mixtures of those solvents, for exampleaqueous solutions, unless otherwise indicated in the description of theprocesses. Such solvent mixtures may also be used in working up, forexample by chromatography or partitioning.

Sulfonimidamides, and their synthesis, are described inChem.Eur.J.2017,23,15189-15193 DOI:10.1002/chem.201703272.

1. A compound of formula (I), or a pharmaceutically acceptable saltthereof:

wherein R, M, W, L, V and T are independently selected from C, CH and N,to form subformulae 1a, 1b, 1c, 1d, 1e and 1f:

A is a linker selected from —C(O)—, —S(O)—, —S(O)₂—, and

Y is N, C or CH; y is 0, 1, 2, 3 or 4; Y

means Y is linked via a single bond to the adjacent carbon atom when Yis CH, or Y is linked via a double bond to the adjacent atom when Y isC, and when Y

is a single bond, Y is carbon unsubstituted or substituted by OH or F;when Y is N, Y

is a single bond; K

means K is linked via a single or double bond to the adjacent atom;wherein: when K

is a double bond, Y

is a single bond, K is CH, J is C, and A is a linker selected from—C(O)—, —S(O)—, —S(O)₂—, and

or when K

is a single bond, K is selected from —CH₂—, —CH₂CH₂—, —NH— and a bond(to form a 5-membered ring:

J is N, and A is a linker selected from —C(O)—, —S(O)—, —S(O)₂—, and

or when K

is a single bond, K is —CH₂—, J is CH, and A is a linker selected from—S(O)—, —S(O)₂—, and

R₅ is independently selected from: —(C₁-C₄)alkyl, —(C₃-C₅)cycloalkyl,and wherein two R₅ substituents on the same ring carbon atom may join,together with the carbon atom to which they are attached, to form a(C₃-C₄)cycloalkyl spiro ring or a 3 or 4-membered heterocyclyl spiroring, wherein said heterocyclyl spiro ring contains ring carbon ringatoms and one ring heteroatom selected from O, N and S, when K

J is a carbon-nitrogen single bond, a R₅ substituent on K and on theadjacent carbon atom may join to form ring C:

wherein ring C is a fused (C₃-C₆)cycloalkyl ring, a fused(C₃-C₆)heterocyclyl ring or a fused phenyl ring, wherein said fused(C₃-C₆)heterocyclyl ring contains ring carbon atoms and one ringheteroatom selected from O, N and S, when K

J is a carbon-carbon single bond, Y is N and Y

is a single bond, and A is a linker selected from —S(O)—, —S(O)₂—, and

a R₅ substituent on K and on the adjacent carbon atom may join to formring C:

and wherein when K is —CH₂— and J is N, two R₅ substituents may join toform a (C₁-C₃)alkylene bridge or a heteroalkylene bridge, wherein saidheteroalkylene bridge is one heteroatom selected from N and O, or is—CH₂—O—CH₂—; and wherein one or more H atoms on the ring:

may be replaced by deuterium; R₁ is: cycloalkenyl, wherein saidcycloalkenyl is a partially unsaturated monocyclic ring containing 5 or6 ring carbon atoms, and said cycloalkenyl is unsubstituted orsubstituted by 1, 2, 3 or 4 R₃₃, wherein R₃₃ is halo, and wherein saidcycloalkenyl or halo-substituted cycloalkenyl is substituted by 0, 1 or2 R₁₅ substituents, or R₁ is heterocyclyl, wherein said heterocyclyl isa 5 or 6 membered fully saturated or partially unsaturated groupcomprising ring carbon atoms and 1 or 2 ring heteroatoms independentlyselected from N, O and S, and wherein said heterocyclyl is unbridged orbridged, and said bridge is 1 or 2 carbon atoms, wherein saidheterocyclyl is unsubstituted or substituted by 1, 2, 3 or 4 R₃₃,wherein R₃₃ is halo, and wherein said heterocyclyl or halo-substitutedheterocyclyl is substituted by 0, 1 or 2 substituents independentlyselected from R₁₅, R₁₆, R₁₇, R₁₈, R₁₉, R₂₀, R₂₂ and R₂₃, or saidheterocyclyl or halo-substituted heterocyclyl is fused to a cyclopropylring, wherein said cyclopropyl ring is unsubstituted or substituted by1, 2 or 3 F, or said heterocyclyl or halo-substituted heterocyclyl has 2substitutents at the same ring carbon atom which join to form acyclopropyl spiro ring, or said heterocyclyl or halo-substitutedheterocyclyl is fused with a (C₃-C₅)heterocycloalkyl ring, wherein said(C₃-C₅)heterocycloalkyl ring contains ring carbon atoms and 1 ring Oatom; or R₁ is heteroaryl, wherein said heteroaryl is a 5 or 6 memberedfully unsaturated monocyclic group comprising ring carbon atoms and 1,2, 3 or 4 ring heteroatoms independently selected from N, O and S,wherein the total number of ring S atoms does not exceed 1, and thetotal number of ring O atoms does not exceed 1, wherein said heteroarylis unsubstituted or substituted by 1, 2 or 3 substituents independentlyselected from R₂₁ and R₃₀, wherein R₂₁ and R₃₀ are independentlyselected from halo and (C₁-C₄)alkyl, wherein said (C₁-C₄)alkyl isunsubstituted or substituted by 1, 2 or 3 halo, or R₁ is phenyl, whereinsaid phenyl is unsubstituted or substituted by 1, 2, 3 or 4 R₃₃, whereinR₃₃ is halo, and wherein said phenyl or halo-substituted phenyl issubstituted by 0, 1 or 2 R₁₅ substituents, or R₁ is (C₂-C₄)alkynyl or(C₂-C₄)alkenyl, wherein said (C₂-C₄)alkynyl and (C₂-C₄)alkenyl areunsubstituted or substituted by (C₁-C₄)alkyl-O—C(O)—, or morpholinyl;each R₁₅, R₁₆, R₁₇, R₁₈, R₁₉, R₂₀, R₂₂ and R₂₃ is independently selectedfrom: halo (C₁-C₄)alkyl-O— unsubstituted or substituted by 1, 2 or 3halo; (C₁-C₄)alkyl unsubstituted or substituted by OH, —O—(C₁-C₂)alkylor 1, 2 or 3 halo, HOC(O)—(CH₂)_(n)—, H₃C—C(O)(CH₂)_(n)—,(C₁-C₄)alkyl-O—C(O)(CH₂)_(n), ═O azetidinyl or pyrrolidinyl, whereinsaid azetidinyl and pyrrolidinyl are linked to the rest of the moleculevia the N atom, and are each unsubstituted or substituted by 1 or 2 F,R₂₅(R₂₄)N—, wherein R₂₄ is H or (C₁-C₄)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo, R₂₅ is H or (C₁-C₄)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo, OH wherein n is 0, 1 or 2, R₂₆ is CH₃, Hor deuterium; R₂₇ is CH₃, H or deuterium; or R₂₆ and R₂₇ join, togetherwith the carbon atom to which they are attached, to form a cyclopropylring; R₂ is the moiety:

R₆ is selected from: H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo, (C₃-C₅)cycloalkyl unsubstituted or substituted by 1,2 or 3 halo, —O—(C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3halo, OH, and CN; R₈ is selected from H, halo, and (C₁-C₄)alkylunsubstituted or substituted by 1, 2 or 3 halo, R₉ is selected from H,O—CH₃, OH, CN, CH₃ and halo; R₂₈ is selected from: SF₅, H, —C(O)H, halo,(C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo,(C₁-C₄)alkynyl, (C₁-C₄)alkenyl, (C₃-C₅)cycloalkyl unsubstituted orsubstituted by 1, 2 or 3 halo, and OCF₃; X is selected from C—R₇ and N,wherein R₇ is H or halo, or R₇ can join, together with R₂₈ or R₆, andthe atoms to which they are attached, to form a fused (C₄-C₆)cycloalkylring, wherein said fused (C₄-C₆)cycloalkyl ring is unsubstituted orsubstituted by 1, 2 or 3 halo, or R₂ is selected from:

wherein R₃₁ is selected from H, halo and CH₃, R₃₂ is selected from H,halo and CH₃, R₃ is selected from: cyclopropyl, O—CH₃, N(CH₃)₂, S—CH₃,and (C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 substituentsindependently selected from halo and OH; R₄ is selected from:

wherein R₁₀, R₁₁, R₁₂, R₁₃ and R₁₄ are independently selected from: H,halo, (C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halosubstituents, (C₁-C₂)alkyl substituted by —O—(C₁-C₂)alkyl or OH,—S—(C₁-C₃)alkyl, —O—(C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or3 halo substituents, OH, (C₃-C₅)cycloalkyl, wherein said(C₃-C₅)cycloalkyl is unsubstituted or substituted by 1 or 2 halo,—O—(C₃-C₅)cycloalkyl, —NR₃₄R₃₅ wherein R₃₄ and R₃₅ are independentlyselected from: H, (C₁-C₄)alkyl, wherein said (C₁-C₄)alkyl isunsubstituted or substituted by OH or —O(C₁-C₂)alkyl, and wherein R₃₄and R₃₅ can join, together with the atom to which they are attached, toform an azetidine, pyrrolidinyl or piperidine ring, wherein saidazetidine, pyrrolidinyl and piperidine are unsubstituted or substitutedwith CH₃; CN, —(C₂-C₄)alkenyl, —(C₂-C₄)alkynyl, —C(O)H, and—C(O)(C₁-C₄)alkyl; and * indicates a point of attachment.
 2. Thecompound of formula (I), or a pharmaceutically acceptable salt thereof,according to claim 1, wherein each R₁ ring atom adjacent to the R₁ ringatom to which said R₁ ring is joined to the remainder of the molecule,is independently unsubstituted or substituted by halo, and said R₁ ringis linked to the remainder of the molecule via a R₁ ring nitrogen atom,or a R₁ ring carbon atom which is double-bonded to an adjacent ringatom.
 3. The compound of formula (I), or a pharmaceutically acceptablesalt thereof, according to claim 1, wherein R₁ is: cycloalkenyl, whereinsaid cycloalkenyl is a partially unsaturated monocyclic ring containing5 or 6 ring carbon atoms, and said cycloalkenyl is unsubstituted orsubstituted by 1, 2, 3 or 4 R₃₃, wherein R₃₃ is halo, and wherein saidcycloalkenyl or halo-substituted cycloalkenyl is substituted by 0, 1 or2 R₁₅ substituents, or R₁ is heterocyclyl, wherein said heterocyclyl isa 5 or 6 membered fully saturated or partially unsaturated groupcomprising ring carbon atoms and 1 or 2 ring heteroatoms independentlyselected from N, O and S, and wherein said heterocyclyl is unbridged orbridged, and said bridge is 1 or 2 carbon atoms, wherein saidheterocyclyl is unsubstituted or substituted by 1, 2, 3 or 4 R₃₃,wherein R₃₃ is halo, and wherein said heterocyclyl or halo-substitutedheterocyclyl is substituted by 0, 1 or 2 substituents independentlyselected from R₁₅, R₁₆, R₁₇, R₁₈, R₁₉, R₂₀, R₂₂ and R₂₃, or R₁ isheteroaryl, wherein said heteroaryl is a 5 or 6 membered fullyunsaturated monocyclic group comprising ring carbon atoms and 1, 2, 3 or4 ring heteroatoms independently selected from N, O and S, wherein thetotal number of ring S atoms does not exceed 1, and the total number ofring O atoms does not exceed 1, wherein said heteroaryl is unsubstitutedor substituted by 1, 2 or 3 substituents independently selected from R₂₁and R₃₀, wherein R₂₁ and R₃₀ are independently selected from halo and(C₁-C₄)alkyl, wherein said (C₁-C₄)alkyl is unsubstituted or substitutedby 1, 2 or 3 halo, and each R₁₅, R₁₆, R₁₇, R₁₈, R₁₉, R₂₀, R₂₂ and R₂₃ isindependently selected from: halo (C₁-C₄)alkyl-O— unsubstituted orsubstituted by 1, 2 or 3 halo; (C₁-C₄)alkyl unsubstituted or substitutedby OH, —O—(C₁-C₂)alkyl or 1, 2 or 3 halo, HOC(O)—(CH₂)_(n)—,H₃C—C(O)(CH₂)_(n)—, (C₁-C₄)alkyl-O—C(O)(CH₂)_(n), ═O azetidinyl orpyrrolidinyl, wherein said azetidinyl and pyrrolidinyl are linked to therest of the molecule via the N atom, and are each unsubstituted orsubstituted by 1 or 2 F, R₂₅(R₂₄)N—, wherein R₂₄ is H or (C₁-C₄)alkylunsubstituted or substituted by 1, 2 or 3 halo, R₂₅ is H or (C₁-C₄)alkylunsubstituted or substituted by 1, 2 or 3 halo, OH wherein n is 0, 1 or2,
 4. The compound of formula (I), or a pharmaceutically acceptable saltthereof, according to claim 1, wherein R₁ is selected from:

R₃₃ is F; R₁₅ is halo, azetidinyl or pyrrolidinyl, wherein saidazetidinyl and pyrrolidinyl are linked to the rest of the molecule viathe N atom, and are unsubstituted or substituted by 1 or 2 F; R₁₆ isR₂₅(R₂₄)N—, wherein R₂₄ is H or (C₁-C₂)alkyl, R₂₅ is H or (C₁-C₂)alkylunsubstituted or substituted by 1, 2 or 3 halo, in particular F; R₁₇ ishalo R₁₈ is halo; R₁₉ is halo; R₂₀ is halo; R₂₁ is (C₁-C₂)alkyl; R₂₂ andR₂₃ are each independently selected from: (C₁-C₄)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo, HOC(O)—(CH₂)_(n)—, H₃C—C(O)(CH₂)_(n)—,(H₃C)₃C—O—C(O)(CH₂)_(n)—; wherein n is 0, 1 or 2; and R₃₀ is CH₃.
 5. Thecompound of formula (I), or a pharmaceutically acceptable salt thereof,according to claim 1, wherein R₁ is selected from:

R₁₅ is F; R₁₆ is R₂₅(R₂₄)N—; R₁₇ is F; R₁₈ is F; R₁₉ is F; R₂₀ is F; R₂₁is CH₃; R₂₂ is CF₃, CHF₂CH₂, HOC(O)—CH₂—, H₃C—C(O)—, (H₃C)₃C—O—C(O)—;R₂₃ is CF₃, CHF₂CH₂—, (H₃C)₃C—O—C(O)—; R₂₄ is CH₃; and R₂₅ is CHF₂CH₂—.6. The compound of formula (I), or a pharmaceutically acceptable saltthereof, according to claim 1, wherein R₁ is selected from:


7. The compound of formula (I), or a pharmaceutically acceptable saltthereof, according to claim 1, wherein R₂ is the moiety:

R₆ is selected from: H, halo, (C₁-C₄)alkyl unsubstituted or substitutedby 1, 2 or 3 halo, (C₃-C₅)cycloalkyl unsubstituted or substituted by 1,2 or 3 halo, —O—(C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3halo, OH, and CN; R₈ is selected from H, halo, and (C₁-C₄)alkylunsubstituted or substituted by 1, 2 or 3 halo, R₉ is selected from H,O—CH₃, OH, CN, CH₃ and halo; R₂₈ is selected from: SF₅, H, —C(O)H, halo,(C₁-C₄)alkyl unsubstituted or substituted by 1, 2 or 3 halo,(C₁-C₄)alkynyl, (C₁-C₄)alkenyl, (C₃-C₅)cycloalkyl unsubstituted orsubstituted by 1, 2 or 3 halo, and OCF₃; and X is selected from C—R₇ andN, wherein R₇ is H or halo.
 8. The compound of formula (I), or apharmaceutically acceptable salt thereof, according to claim 1, whereinR₂ is the moiety:

R₆ is selected from H, Cl, CH₃, F and Br; R₈ is selected from H, Cl, Fand CF₃; R₉ is selected from H, CH₃ and Cl; R₂₈ is selected from CF₃,CF₂H, —CH₂CH₃, Cl, SF₅, Br and —C(O)H; X is selected from C—R₇ and N;and R₇ is selected from H and F.
 9. The compound of formula (I), or apharmaceutically acceptable salt thereof, according to claim 1, whereinR₂ is selected from:


10. The compound of formula (I), or a pharmaceutically acceptable saltthereof, according to claim 1, wherein R₂₆ is H and R₂₇ is H.
 11. Thecompound of formula (I), or a pharmaceutically acceptable salt thereof,according to claim 1, wherein R₃ is (C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 substituents independently selected from haloand OH.
 12. The compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to claim 1, wherein Y is N and Y

is Y linked by a single bond.
 13. The compound of formula (I), or apharmaceutically acceptable salt thereof, according to claim 1, whereinK

is K linked by a single bond, and K is selected from —CH₂—, —CH₂CH₂—,—NH— and a bond (to form a 5-membered ring:

J is N, and A is a linker selected from —C(O)—, —S(O)—, —S(O)₂—, and


14. The compound of formula (I), or a pharmaceutically acceptable saltthereof, according to claim 1, wherein A is a linker selected from—C(O)— and —S(O)₂.
 15. The compound of formula (I), or apharmaceutically acceptable salt thereof, according to claim 1, whereinR₅ is independently selected from: —(C₁-C₄)alkyl, preferably methyl, andwherein two R₅ substituents on the same ring carbon atom may join,together with the carbon atom to which they are attached, to form a(C₃-C₄)cycloalkyl spiro ring or a 3 or 4-membered heterocyclyl spiroring, wherein said heterocyclyl spiro ring contains ring carbon ringatoms and one ring heteroatom selected from O, N and S, when K

J is a carbon-nitrogen single bond, a R₅ substituent on K and on theadjacent carbon atom may join to form ring C:

wherein ring C is a fused (C₃-C₆)cycloalkyl ring, in particular a fusedcyclobutyl ring, a fused (C₃-C₆)heterocyclyl ring or a fused phenylring, wherein said fused (C₃-C₆)heterocyclyl ring contains ring carbonatoms and one ring heteroatom selected from O, N and S, and wherein whenK is —CH₂— and J is N, two R₅ substituents may join to form a(C₁-C₃)alkylene bridge or a heteroalkylene bridge, wherein saidheteroalkylene bridge is one heteroatom selected from N and O, or is—CH₂—O—CH₂—.
 16. The compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to claim 1, wherein R₅ isindependently selected from: CH₃, and y is 1 or 2, and when K

J is a carbon-nitrogen single bond, a R₅ substituent on K and on theadjacent carbon atom may join to form ring C:

wherein ring C is a fused cyclobutyl ring.
 17. The compound of formula(I), or a pharmaceutically acceptable salt thereof, according to claim1, wherein y is 0, 1, or
 2. 18. The compound of formula (I), or apharmaceutically acceptable salt thereof, according to claim 1, whereinthe moiety:

is selected from a)


19. The compound of formula (I), or a pharmaceutically acceptable saltthereof, according to claim 1, wherein R₄ is selected from:

wherein R₁₀ is selected from H, halo, (C₁-C₂)alkyl unsubstituted orsubstituted by 1, 2 or 3 halo substituents, —O—(C₁-C₂)alkylunsubstituted or substituted by 1, 2 or 3 halo substituents; R₁₁ isselected from H, halo, (C₁-C₂)alkyl unsubstituted or substituted by 1, 2or 3 halo substituents; R₁₂ is selected from H, halo, (C₁-C₂)alkylunsubstituted or substituted by 1, 2 or 3 halo substituents; R₁₃ isselected from H, —S—CH₃, halo, (C₁-C₂)alkyl unsubstituted or substitutedby 1, 2 or 3 halo substituents; and R₁₄ is selected from H, halo,(C₁-C₂)alkyl unsubstituted or substituted by 1, 2 or 3 halosubstituents, O—(C₁-C₂)alkyl unsubstituted or substituted by 1, 2 or 3halo substituents, and cyclopropyl.
 20. The compound of formula (I), ora pharmaceutically acceptable salt thereof, according to claim 1,wherein R₄ is selected from:

wherein R₁₀ is selected from H, F, Cl, CH₃ and OCF₃; R₁₁ is selectedfrom H, Cl, F, and CH₃; R₁₂ is selected from H, Cl and CH₃; R₁₃ isselected from H, —S—CH₃ and CH₃; and R₁₄ is selected from H, CH₃,—CH₂CH₃, cyclopropyl, —OCHF₂, OCF₃ and Cl.
 21. The compound of formula(I), or a pharmaceutically acceptable salt thereof, according to claim1, wherein R₄ is selected from:


22. The compound of formula (I), or a pharmaceutically acceptable saltthereof, according to claim 1, wherein formula (I) is formula 1a:


23. The compound of formula (I), or a pharmaceutically acceptable saltthereof, according to claim 1, wherein formula (I) is formula 1g:

and Y is N or CH, in particular N.
 24. The compound of formula (I), or apharmaceutically acceptable salt thereof, according to claim 1, whereinformula (I) is selected from


25. A compound of formula (I), or a pharmaceutically acceptable saltthereof, according to claim 1, wherein the compound is selected from:


26. A compound of formula (I), or a pharmaceutically acceptable saltthereof, according to claim 1, wherein the compound is selected from:


27. A compound of formula (I), or a pharmaceutically acceptable saltthereof, according to claim 1, wherein the compound is:


28. (canceled)
 29. (canceled)
 30. A compound of formula (I), or apharmaceutically acceptable salt thereof, according to claim 1, whereinthe compound is in non-zwitterionic form.
 31. A compound of formula (I),or a pharmaceutically acceptable salt thereof, according to claim 1,wherein the compound is in zwitterionic form. 32-34. (canceled)
 35. Acompound of formula (I), or a pharmaceutically acceptable salt thereof,according to claim 1, wherein the compound is a sodium salt
 36. Acompound of formula (I), or a pharmaceutically acceptable salt thereof,according to claim 1, wherein the compound is in amorphous form.
 37. Acompound of formula (I), or a pharmaceutically acceptable salt thereof,according to claim 1, in crystalline form.
 38. (canceled)
 39. Acombination comprising a compound of formula (I), or a pharmaceuticallyacceptable salt thereof, according to claim 1, and one or moreadditional therapeutically active agents. 40-45. (canceled)
 46. Apharmaceutical composition comprising a compound of formula (I) or apharmaceutically acceptable salt thereof, according to claim 1, and oneor more pharmaceutically acceptable carriers. 47-53. (canceled)
 54. Amethod of modulating WRN activity in a subject, wherein the methodcomprises administering to the subject a therapeutically effectiveamount of the compound of formula (I), or a pharmaceutically acceptablesalt thereof, according to claim
 1. 55. A method of inhibiting WRN in asubject, wherein the method comprises administering to the subject atherapeutically effective amount of the compound of formula (I), or apharmaceutically acceptable salt thereof, according to claim
 1. 56. Amethod of treating a disorder or disease which can be treated by WRNinhibition in a subject, comprising administering to the subject atherapeutically effective amount of the compound of formula (I), or apharmaceutically acceptable salt thereof, according to claim
 1. 57. Amethod of treating cancer in a subject, comprising administering to thesubject a therapeutically effective amount of the compound of formula(I), or a pharmaceutically acceptable salt thereof, according toclaim
 1. 58-66. (canceled)
 67. An intermediate compound selected from: acompound, or salt thereof, of formula A:

wherein R₁, R₂, R₃, R₂₆, R₂₇, R₅, y and Y are as defined in claim 1, acompound:

intermediate AK, a compound, or salt thereof, of Formula B:

wherein R₁, R₂, R₃, R₂₆, R₂₇, R₅, y and Y are as defined in claim 1, andPG₁ is a protecting group, a compound, or salt thereof, which is:

Intermediate AK step 1 product. a compound, or salt thereof, of FormulaC:

wherein R₁, R₃, R₅, y and Y are as defined in claim 1, and PG₂ is aprotecting group, a compound, or salt thereof, which is:

Intermediate AF a compound, or salt thereof, or a tautomer thereof, ofFormula D:

wherein R₃, R₅, y and Y are as defined in claim 1, and PG₃ is aprotecting group, a compound, or salt thereof, which is:

intermediate AF step 1 product, and a compound, or salt thereof, whichis:

Intermediate EH.